Difference between revisions of "Team:Munich/LabJournal"

Line 78: Line 78:
 
<div class="month" id="March" >
 
<div class="month" id="March" >
 
<div class="date firstOfMonth">Tuesday 21st</div>
 
<div class="date firstOfMonth">Tuesday 21st</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>Read-out</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Wednesday 22nd</div>
 
    <div class="date">Wednesday 22nd</div>
−
     
+
    <ul class="myEntry cas13a">
 +
    <li class="myTitel">
 +
    <b>Cas13a41234</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Thursday 23rd</div>
 
    <div class="date">Thursday 23rd</div>
−
     
+
    <ul class="myEntry target">
 +
    <li class="myTitel">
 +
    <b>Target</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Friday 24th</div>
 
    <div class="date">Friday 24th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Monday 27th</div>
 
    <div class="date">Monday 27th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Tuesday 28th</div>
 
    <div class="date">Tuesday 28th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Wednesday 29th</div>
 
    <div class="date">Wednesday 29th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Thursday 30th</div>
 
    <div class="date">Thursday 30th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Friday 31rd</div>
 
    <div class="date">Friday 31rd</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
</div>
 
</div>
 
<div class="month" id="April" >
 
<div class="month" id="April" >
 
<div class="date firstOfMonth">Monday 3rd</div>
 
<div class="date firstOfMonth">Monday 3rd</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>Read-out</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Tuesday 4th</div>
 
    <div class="date">Tuesday 4th</div>
−
     
+
    <ul class="myEntry cas13a">
 +
    <li class="myTitel">
 +
    <b>Cas13a41234</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Wednesday 5th</div>
 
    <div class="date">Wednesday 5th</div>
−
     
+
    <ul class="myEntry target">
 +
    <li class="myTitel">
 +
    <b>Target</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Thursday 6th</div>
 
    <div class="date">Thursday 6th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Friday 7th</div>
 
    <div class="date">Friday 7th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Monday 10th</div>
 
    <div class="date">Monday 10th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Tuesday 11th</div>
 
    <div class="date">Tuesday 11th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Wednesday 12th</div>
 
    <div class="date">Wednesday 12th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Thursday 13th</div>
 
    <div class="date">Thursday 13th</div>
−
    <div class="date">Friday 14th</div>
+
    <ul class="myEntry readout">
−
     
+
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul><div class="date">Friday 14th</div>
 +
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Monday 17th</div>
 
    <div class="date">Monday 17th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Tuesday 18th</div>
 
    <div class="date">Tuesday 18th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Wednesday 19th</div>
 
    <div class="date">Wednesday 19th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date ">Thursday 20th</div>
 
    <div class="date ">Thursday 20th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Friday 21st</div>
 
    <div class="date">Friday 21st</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Monday 24th</div>
 
    <div class="date">Monday 24th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Tuesday 25th</div>
 
    <div class="date">Tuesday 25th</div>
 
<ul class="myEntry cas13a">       
 
<ul class="myEntry cas13a">       
Line 250: Line 750:
  
 
    <div class="date">Friday 28th</div>
 
    <div class="date">Friday 28th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
</div>
 
</div>
 
                       <div class="month" id="May" >
 
                       <div class="month" id="May" >
 
    <div class="date firstOfMonth">Monday 1st</div>
 
    <div class="date firstOfMonth">Monday 1st</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Tuesday 2nd</div>
 
    <div class="date">Tuesday 2nd</div>
 
<ul class="myEntry cas13a"> 
 
<ul class="myEntry cas13a"> 
Line 661: Line 1,201:
 
    </li>
 
    </li>
 
   </ul>     <div class="date">Wednesday 17th</div>
 
   </ul>     <div class="date">Wednesday 17th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Thursday 18th</div>
 
    <div class="date">Thursday 18th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Friday 19th</div>
 
    <div class="date">Friday 19th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Monday 22nd</div>
 
    <div class="date">Monday 22nd</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Tuesday 23rd</div>
 
    <div class="date">Tuesday 23rd</div>
 
<ul class="myEntry cas13a"> 
 
<ul class="myEntry cas13a"> 
Line 1,182: Line 1,802:
 
    </li>
 
    </li>
 
    </ul>
 
    </ul>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Monday 5th</div>
 
    <div class="date">Monday 5th</div>
 
<ul class="myEntry cas13a"> 
 
<ul class="myEntry cas13a"> 
Line 1,658: Line 2,298:
 
</ul>
 
</ul>
 
    <div class="date">Thursday 15th</div>
 
    <div class="date">Thursday 15th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Friday 16th</div>
 
    <div class="date">Friday 16th</div>
 
<ul class="myEntry cas13a"> 
 
<ul class="myEntry cas13a"> 
Line 1,931: Line 2,591:
 
</ul>
 
</ul>
 
    <div class="date">Thursday 22nd</div>
 
    <div class="date">Thursday 22nd</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Friday 23rd</div>
 
    <div class="date">Friday 23rd</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
                             <ul class="myEntry other">
 
                             <ul class="myEntry other">
 
    <li class="myTitel">
 
    <li class="myTitel">
Line 1,967: Line 2,667:
 
    </ul>
 
    </ul>
 
    <div class="date">Monday 26th</div>
 
    <div class="date">Monday 26th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <ul class="myEntry other">
 
    <ul class="myEntry other">
 
    <li class="myTitel">
 
    <li class="myTitel">
Line 2,427: Line 3,147:
  
 
    <div class="date">Thursday 6th</div>
 
    <div class="date">Thursday 6th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Friday 7th</div>
 
    <div class="date">Friday 7th</div>
 
  <ul class="myEntry cas13a"> 
 
  <ul class="myEntry cas13a"> 
Line 2,529: Line 3,269:
  
 
</ul>     <div class="date">Thursday 13th</div>
 
</ul>     <div class="date">Thursday 13th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Friday 14th</div>
 
    <div class="date">Friday 14th</div>
 
<ul class="myEntry cas13a"> 
 
<ul class="myEntry cas13a"> 
Line 2,592: Line 3,352:
 
    </li>
 
    </li>
 
    </ul>
 
    </ul>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Tuesday 18th</div>
 
    <div class="date">Tuesday 18th</div>
 
<ul class="myEntry cas13a"> 
 
<ul class="myEntry cas13a"> 
Line 2,807: Line 3,587:
 
</ul>
 
</ul>
 
    <div class="date">Monday 31th</div>
 
    <div class="date">Monday 31th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
                     </div>
 
                     </div>
 
<div class="month" id="August" >
 
<div class="month" id="August" >
 
    <div class="date firstOfMonth">Tuesday 1st</div>
 
    <div class="date firstOfMonth">Tuesday 1st</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Wednesday 2nd</div>
 
    <div class="date">Wednesday 2nd</div>
 
    <ul class="myEntry other">
 
    <ul class="myEntry other">
Line 2,847: Line 3,667:
 
    </li>
 
    </li>
 
    </ul>
 
    </ul>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Thursday 3rd</div>
 
    <div class="date">Thursday 3rd</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Friday 4th</div>
 
    <div class="date">Friday 4th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Monday 7th</div>
 
    <div class="date">Monday 7th</div>
 
    <ul class="myEntry other">
 
    <ul class="myEntry other">
Line 2,878: Line 3,758:
 
    </li>
 
    </li>
 
    </ul>
 
    </ul>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Tuesday 8th</div>
 
    <div class="date">Tuesday 8th</div>
 
    <ul class="myEntry other">
 
    <ul class="myEntry other">
Line 2,907: Line 3,807:
 
    </li>
 
    </li>
 
    </ul>
 
    </ul>
−
                              
+
                             <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Wednesday 9th</div>
 
    <div class="date">Wednesday 9th</div>
 
    <ul class="myEntry other">
 
    <ul class="myEntry other">
Line 2,943: Line 3,863:
 
    </li>
 
    </li>
 
    </ul>    
 
    </ul>    
−
                              
+
                             <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Thursday 10th</div>
 
    <div class="date">Thursday 10th</div>
 
    <ul class="myEntry other">
 
    <ul class="myEntry other">
Line 2,966: Line 3,906:
 
    </li>
 
    </li>
 
    </ul>    
 
    </ul>    
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Friday 11th</div>
 
    <div class="date">Friday 11th</div>
 
    <ul class="myEntry other">
 
    <ul class="myEntry other">
Line 3,004: Line 3,964:
 
    </li>
 
    </li>
 
    </ul>    
 
    </ul>    
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Monday 14th</div>
 
    <div class="date">Monday 14th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Tuesday 15th</div>
 
    <div class="date">Tuesday 15th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Wednesday 16th</div>
 
    <div class="date">Wednesday 16th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Thursday 17th</div>
 
    <div class="date">Thursday 17th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Friday 18th</div>
 
    <div class="date">Friday 18th</div>
 
                             <div class="date">Sunday 20th</div>
 
                             <div class="date">Sunday 20th</div>
Line 3,572: Line 4,632:
 
    </li>
 
    </li>
 
    </ul>
 
    </ul>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Thursday 31st</div>
 
    <div class="date">Thursday 31st</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <ul class="myEntry other">
 
    <ul class="myEntry other">
 
    <li class="myTitel">
 
    <li class="myTitel">
Line 3,795: Line 4,895:
  
 
    <div class="date">Wednesday 6th</div>
 
    <div class="date">Wednesday 6th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Thursday 7th</div>
 
    <div class="date">Thursday 7th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Friday 8th</div>
 
    <div class="date">Friday 8th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <ul class="myEntry other">
 
    <ul class="myEntry other">
 
    <li class="myTitel">
 
    <li class="myTitel">
Line 3,832: Line 4,992:
  
 
    <div class="date">Monday 11th</div>
 
    <div class="date">Monday 11th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Tuesday 12th</div>
 
    <div class="date">Tuesday 12th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Wednesday 13th</div>
 
    <div class="date">Wednesday 13th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
        <ul class="myEntry other">
 
        <ul class="myEntry other">
 
    <li class="myTitel">
 
    <li class="myTitel">
Line 3,956: Line 5,176:
 
    </ul>
 
    </ul>
 
    <div class="date">Monday 18th</div>
 
    <div class="date">Monday 18th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Tuesday 19th</div>
 
    <div class="date">Tuesday 19th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
  
  
Line 3,987: Line 5,247:
 
    </ul>
 
    </ul>
 
    <div class="date">Wednesday 20th</div>
 
    <div class="date">Wednesday 20th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Thurdsay 21st</div>
 
    <div class="date">Thurdsay 21st</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Friday 22nd</div>
 
    <div class="date">Friday 22nd</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Monday 25th</div>
 
    <div class="date">Monday 25th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Tuesday 26th</div>
 
    <div class="date">Tuesday 26th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Wednesday 27th</div>
 
    <div class="date">Wednesday 27th</div>
 
    <ul class="myEntry other">
 
    <ul class="myEntry other">
Line 4,020: Line 5,380:
 
    </li>
 
    </li>
 
    </ul>
 
    </ul>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Thursday 28th</div>
 
    <div class="date">Thursday 28th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Friday 29th</div>
 
    <div class="date">Friday 29th</div>
 
    <ul class="myEntry other">
 
    <ul class="myEntry other">
Line 4,119: Line 5,519:
 
<div class="month" id="October" >
 
<div class="month" id="October" >
 
    <div class="date fistOfMonth">Monday 2nd</div>
 
    <div class="date fistOfMonth">Monday 2nd</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Tuesday 3rd</div>
 
    <div class="date">Tuesday 3rd</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Wednesday 4th</div>
 
    <div class="date">Wednesday 4th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Thursday 5th</div>
 
    <div class="date">Thursday 5th</div>
 
     
 
     
  
 
    <div class="date">Friday 6th</div>
 
    <div class="date">Friday 6th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Monday 9th</div>
 
    <div class="date">Monday 9th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Wednesday 11th</div>
 
    <div class="date">Wednesday 11th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Thursday 12th</div>
 
    <div class="date">Thursday 12th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Friday 13th</div>
 
    <div class="date">Friday 13th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Monday 16th</div>
 
    <div class="date">Monday 16th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Tuesday 17th</div>
 
    <div class="date">Tuesday 17th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Wednesday 18th</div>
 
    <div class="date">Wednesday 18th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Thursday 19th</div>
 
    <div class="date">Thursday 19th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Friday 20th</div>
 
    <div class="date">Friday 20th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Monday 23rd</div>
 
    <div class="date">Monday 23rd</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Tuesday 24th</div>
 
    <div class="date">Tuesday 24th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Wednesday 25th</div>
 
    <div class="date">Wednesday 25th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Thursday 26th</div>
 
    <div class="date">Thursday 26th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Friday 27th</div>
 
    <div class="date">Friday 27th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Monday 30th</div>
 
    <div class="date">Monday 30th</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
    <div class="date">Tuesday 31st</div>
 
    <div class="date">Tuesday 31st</div>
−
     
+
    <ul class="myEntry readout">
 +
    <li class="myTitel">
 +
    <b>readout</b>
 +
    </li>
 +
    <li class="myProtocol">
 +
    Protocol: <a href="#">Protocol</a>
 +
    </li>
 +
    <li class="myParticipants">
 +
    Participants:
 +
    </li>
 +
    <li class="myObservations">
 +
    Observations
 +
    <ul>
 +
    <li></li>
 +
    <li></li>
 +
    </ul>
 +
    </li>
 +
    <li class="myResults">
 +
    Results
 +
    </li>
 +
    </ul>
 
                   </div>
 
                   </div>
 
</div>
 
</div>

Revision as of 12:41, 26 October 2017

Description

Design

Demonstrate

Measurement

Applied Design

Final Results

Achievements

Protocols

Notebook

Safety

Parts

Improved Part

Part Collection

InterLab

Model

Software

Collaboration

Detector

Quake Valve

Paperstrips

Sample Processing

Silver

Gold

Engagement

Collaborations

Team members

Sponsors

Attributions

Gallery

Labbook

According to all known laws of aviation, there is no way a bee should be able to fly. Its wings are too small to get its fat little body off the ground. The bee, of course, flies anyway because bees don't care what humans think is impossible. Yellow, black. Yellow, black. Yellow, black. Yellow, black.

Tuesday 21st
  • Read-out
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Wednesday 22nd
  • Cas13a41234
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Thursday 23rd
  • Target
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Friday 24th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Monday 27th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Tuesday 28th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Wednesday 29th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Thursday 30th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Friday 31rd
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Monday 3rd
  • Read-out
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Tuesday 4th
  • Cas13a41234
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Wednesday 5th
  • Target
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Thursday 6th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Friday 7th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Monday 10th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Tuesday 11th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Wednesday 12th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Thursday 13th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Friday 14th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Monday 17th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Tuesday 18th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Wednesday 19th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Thursday 20th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Friday 21st
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Monday 24th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Tuesday 25th
  • Plating of DH5α p2CT-His-MBP-Lbu-Cas13a-WT and DH5α p2CT-His-MBP-Lsh-Cas13a-WT on LBCarb agar plates
  • Protocol: -
  • Participants: Ludwig
  • Observations:
    • Incubation at 37 °C over night.

  • Results

    Bacteria grew and single colonies were visible.
Wednesday 26th
  • Preparation of 6 ml LBCarb cultures of DH5α p2CT-His-MBP-Lbu-Cas13a-WT and DH5α p2CT-His-MBP-Lsh-Cas13a-WT on LBCarb agar plates
  • Protocol: -
  • Participants: Ludwig
  • Observations:
    • Incubation at 37 °C over night.

  • Results

    Bacteria cultures grew.
Thursday 27th
  • Preparation of cryo stocks for DH5α p2CT-His-MBP-Lbu-Cas13a-WT and DH5α p2CT-His-MBP-Lsh-Cas13a-WT
  • Protocol: Cryo stocks
  • Participants: Ludwig
  • Minipreparation of DH5α Cas13a overnight cultures
  • Protocol: Minipreparation
  • Participants: Ludwig
  • Electro transformation of p2CT-His-MBP-Lbu-Cas13a-WT and p2CT-His-MBP-Lsh-Cas13a-WT in E. coli Rosetta.
  • Protocol: Electro transformation
  • Participants: Ludwig
  • Observations:
    • 1 µl of plasmid DNA was used.
    • Bacteria were plated on LBCarb plates without additional Cm.

  • Results

    Bacteria lost second plasmid probably.
Friday 28th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Monday 1st
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Tuesday 2nd
  • Electro transformation of p2CT-His-MBP-Lbu-Cas13a-WT and p2CT-His-MBP-Lsh-Cas13a-WT in E. coli Rosetta.
  • Protocol: Electro transformation
  • Participants: Ludwig
  • Observations:
    • 1 µl of plasmid DNA was used.
    • Bacteria were plated on LBCarb + Cm plates

  • Results

    Bacteria grew and single colonies were visible.
  • Preparations of buffers for Cas13a purification.
  • Protocol: Protein purification
  • Participants: Chris, Ludwig, Julian
  • Observations:
    • TCEP changes the pH. Readjustement after adding neccessary.
  • Electro transformation of p2CT-His-MBP-Lbu-Cas13a-WT and p2CT-His-MBP-Lsh-Cas13a-WT in E. coli Rosetta.
  • Protocol: Electro transformation
  • Participants: Ludwig
  • Observations:
    • 1 µl of plasmid DNA was used.
    • Bacteria were plated on LBCarb + Cm plates

  • Results

    Bacteria grew, but only few single colonies were visible.
Wednesday 3rd
  • Electro transformation of p2CT-His-MBP-Lbu-Cas13a-WT and p2CT-His-MBP-Lsh-Cas13a-WT in E. coli Rosetta.
  • Protocol: Electro transformation
  • Participants: Ludwig, Chris
  • Observations:
    • 1 µl of plasmid DNA was used.
    • Bacteria were plated on LBCarb + Cm plates

  • Results

    This time bacteria grew well and a lot of single colonies were visible.
  • Preparation of 25 ml LBCarb + Cm cultures of Rosetta p2CT-His-MBP-Lbu-Cas13a-WT and Rosetta p2CT-His-MBP-Lsh-Cas13a-WT and incubated at 37 °C overnight.
  • Protocol: Protein Expression
  • Participants: Ludwig
Thursday 4th
  • Harvesting of Rosetta cells with overexpressed Cas13a Lbu and Cas13a Lsh
  • Protocol: Protein expression
  • Participants: Ludwig, Chris

  • Results

    Bacterial pellets were stored at -80 °C.
Friday 5th
  • Protein purification of Cas13a Lbu
  • Protocol: Äkta protein purification
  • Participants: Ludwig, Chris
  • Observations:
    • One test sample was run on the Äkta before the actual run. Only the second run was analyzed with a SDS PAGE.

  • Results

    xxxChange xxxChange xxxChange
Monday 8th
  • Transformation of TEV plasmid (form lab in the chemistry department) in DH5α
  • Protocol: Electro Transformation
  • Participants: Sven
Tuesday 9th
  • Dialysis of pooled peak fractions of Cas13a Lbu and Cas13a Lsh into ion exchange buffer
  • Protocol: Äkta protein purification
  • Participants: Ludwig, Chris, Sven
  • PCR of NT and T crDNA (for Cas13a Lbu and Cas13a Lsh)
  • Protocol:
  • Participants: Sven
  • Observations:
    • PCR didn't work.
Wednesday 10th
  • Test of different lysis methods for total RNA extraction with Norgen kit
  • Protocol: Total RNA Purification with Norgen Kit
  • Participants: Julian, Milica, Jorge
  • Observations:
    • Three lysis methods were used: sonifcation, lysozyme digestion and heat lysis in SDS.
    • Lysozyme digestion: According to protocol.
    • Heat lysis: Sample was incubated in 2% SDS at 75 °C for 15 min.

  • Results

    See results from "Phenol/Chloroform purification of RNA from in-vitro transcription" from 11.05.17 for gel picture.
  • Upconcentration fo Cas13a Lbu and Cas13 Lsh after dialysis
  • Protocol: Äkta protein purification
  • Participants: Ludwig, Chris

  • Results

Thursday 11th
  • Test of different lysis methods for Phenol/Chloroform total RNA extraction.
  • Protocol: Phenol/Chloroform purification of RNA
  • Participants: Milica, Sven
  • Observations:
    • Three lysis method were used: sonification, lysozyme digestion and heat lysis in SDS.
    • Lysozyme digestion: according to the total RNA purification with Norgen Kit protocol.
    • Heat lysis: Sample was incubated in 2% SDS at 75 °C for 15 min.
    • The samples were incubated at -80 °C overnight and the purification finished the next day.

  • Results

    See results from "Phenol/Chloroform purification of RNA from in-vitro transcription" from 11.05.17.
  • Phenol/Chloroform purification of RNA from in-vitro transcription
  • Protocol: Phenol/Chloroform purification of RNA
  • Participants: Dawafuti, Sven
  • Observations
    • The samples were incubated at -80 °C overnight and the purification finished the next day.

  • Results

Friday 12th
  • Urea PAGE of in vitro transcription samples
  • Protocol: UREA PAGE
  • Participants: Sven

  • Results

Monday 15th
  • Preparation fo buffers for Heparin purification of Cas13a Lbu and Cas13a Lsh
  • Protocol: Äkta protein purification
  • Participants: Ludwig, Chris
Tuesday 16th
  • Heparin purification of Cas13a Lbu and Cas13 Lsh
  • Protocol: Äkta protein purification
  • Participants: Ludwig, Dawa

  • Results

    xxxChange
Wednesday 17th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Thursday 18th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Friday 19th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Monday 22nd
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Tuesday 23rd
  • PCR of Cas13a Lwa part1 and part2 and backbone pSB1C3_ccdB
  • Protocol: PCR
  • Participants: Ludwig, Chris, Rob
  • Observations:
    • Q5 polymerase was used
    • Primers Lwa part1 and part2: VF and VR
    • Primers pSB1C3_ccdB: p-pSB-GG-Fwd, p-pSB-GG-Rev

  • Results

    xxxChange
  • Assembly of Cas13a Lwa part1 and part2 with pSB1C3_ccdB using GoldenGate Cloning
  • Protocol: GoldenGate Assembly
  • Participants: Ludwig, Chris, Rob
  • Observations:
    • Insert to vector ratio: 2:1 and 3:1
  • Electro Transformation of assembled Lwa parts and pSB1C3 into DH5α and plating on LBCm agar plates
  • Protocol: Electro Transformation
  • Participants: Ludwig, Chris, Rob
  • Observations:
    • Next day: Only a few colonies.
  • Electro Transformation of TEV biobricks (pSB1C3-BBa_K1319008 and pSB1C3-BBa_K1319004) and plating on LBCm agar plates
  • Protocol: Electro Transformation
  • Participants: Ludwig, Chris, Rob
Wednesday 24th
  • Repeat: Assembly of Cas13a Lwa part1 and part2 with pSB1C3_ccdB using GoldenGate Cloning
  • Protocol: GoldenGate Assembly
  • Participants: Ludwig, Erika, Dawa, Chris
  • Observations:
    • Insert to vector ratio: 2:1 and 3:1
  • Electro Transformation of assembled Lwa parts and pSB1C3 into DH5α and plating on LBCm agar plates
  • Protocol: Electro Transformation
  • Participants: Ludwig, Erika, Dawa, Chris
  • Repeat: PCR of Cas13a Lwa part1 and part2 and backbone pSB1C3_ccdB
  • Protocol: PCR
  • Participants: Ludwig, Erika, Dawa, Chris
  • Observations:
    • Q5 polymerase was used
    • Primers Lwa part1 and part2: VF and VR
    • Primers pSB1C3_ccdB: p-pSB-GG-Fwd, p-pSB-GG-Rev

  • Results

    xxxChange
  • Preparation of 5 ml LBCm overnight culture of one picked DH5α pSB1C3-Cas13a-LwaLwa colony (from first GoldenGate assembly)
  • Protocol: -
  • Participants: Ludwig, Erika, Dawa, Chris
Thursday 25th
  • Miniprep of DH5α pSB1C3-Cas13a-Lwa overnight culture
  • Protocol: Minipreparation
  • Participants: Dawa, Chris
  • Observations:
    • pSB1C3-Cas13a-Lwa was then used for an analytical restriction digest and was sent for sequencing (with primer VR)
  • Analytical restriction digest with pSB1C3-Cas13a-LwaLwa with XbaI (single digest) and XbaI + SpeI (double digest)
  • Protocol: Restriction digest
  • Participants: Dawa, Chris

  • Results

  • Repeat: PCR of Lwa part2 and also GoldenGate was redone with new amplified Lwa part2
  • Protocol: PCR and GoldenGate
  • Participants:
  • Observations:
    • PCR Primers: VF, VR
    • Insert:Vecor 3:1 (GoldenGate)

  • Results

Friday 26th
  • Transformation of 1 µl GoldenGate sample in DH5α and plating on LBCm agar plates
  • Protocol: Transformation
  • Participants: Ludwig, Dawa, Chris
  • Observations:
    • No colonies visible
  • PCR of BBa_K1319004 and BBa_K1319008 (TEV) to check if biobricks are in plasmid
  • Protocol: PCR
  • Participants: Ludwig, Dawa, Chris

  • Results

  • Electro transformation of BBa_K1319004 and BBa_K1319008 (TEV) in DH5α and plating on LBCm agar plates
  • Protocol:
  • Participants:
  • Observations:
    • No colonies for BBa_K1319008
Monday 29th
  • Chemical transformation of BBa_K1319008 (TEV) in DH5α and plating on LBCm agar plates
  • Protocol: Chemical Transformation
  • Participants: Ludwig, Dawa, Erika
  • Observations:
    • 3 µl (= 750 pg) were used
  • Chemical transformation of GolenGate sample from 24.05. and also from 26.05. in DH5α and plating on LBCm agar plates
  • Protocol: Chemical Transformation
  • Participants: Ludwig, Dawa, Erika
  • Observations:
    • 5 µl for each transformation were used
Tuesday 30th
  • Colony PCR of DH5α pSB1C3-Cas13a-LwaLwa (from GoldenGate)
  • Protocol: cPCR
  • Participants: Chris, Dawa

  • Results

    xxxChange
  • Colony PCR of DH5α pSB1C3-BBa_K1319004 and pSB1C3-BBa_K1319008 (TEV)
  • Protocol: cPCR
  • Participants: Chris, Dawa
  • Observations:
    • Primers: VF, VR
    • Overnight cultures from cultures that showed right cPCR bands were set up

  • Results

    xxxChange
Wednesday 31st
  • Preparation of buffers for Cas13a purification (Elution, Wash, Ion exchange and SEC)
  • Protocol: Äkta purification
  • Participants: Milica, Ludwig, Flo, Sven, Jorge
  • Observations:
  • Minipreparation of DH5α pSB1C3-BBa_K1319004 and pSB1C3-BBa_K1319008 (TEV) overnight cultures
  • Protocol: Miniprep
  • Participants: Milica, Ludwig, Flo, Sven, Jorge
  • Observations:
    • Each construct was sent for sequencing with primers VR and VF
Thursday 1st
  • Colony PCR of DH5α pSB1C3-Cas13a-Lwa
  • Protocol: cPCR
  • Participants: Ludwig, Erika, Sven
  • Observations:
    • Primers: VR, VF

  • Results

    xxxChange
  • Äkta purification of Cas13a Lbu and Cas13a Lwa (Affinity chromatography)
  • Protocol: Äkta protein purification
  • Participants: Ludwig, Erika, Sven

  • Results

    xxxChange
Friday 2nd
  • Total RNA extraction of E. coli W3110 with Norgen kit
  • Protocol: Total RNA Purification with Norgen Kit
  • Participants: Jorge
  • Observations:
    • Instead of using TE and lysozyme to lyse the cells, they were resuspenden in 2% SDS in TAE and incubated at 75 ºC for 15 min.
    • The 6 samples were stored at -80 ºC and labeled T RNA #1-6
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Monday 5th
  • Preparation of overnight cultures from Rosetta pSB1C3-His-MBP-Lbu-Cas13a-WT and Rosetta p2CT-His-MBP-Lsh-Cas13a-WT glycerol stocks, DH5α pSB-Cas13a-Lwa cPCR colony 5 and DH5α pSB1C3-BBa_K1319004 cPCR colony 1 and DH5α pSB1C3-BBa_K1319008 cPCR colony 1
  • Protocol: -
  • Participants: Sven
  • RNA Adsorption Test on first 3D print
  • Participants: Jorge
  • Notes:
    • Total RNA sample 5 used, prepared on Friday 02/06 by Jorge (Concentration: 126 ng/µl)
    • Procedure:
      • Take 1 µl presample from stock ; add with 2 µl nf H2O and 3 µl 2x RNA LD
      • Put 5 µl of sample on device
      • Take 1 µl of sample immediately, add with 2 µl nf H2O and 3 µl 2x RNA LD
      • Wait 15 minutes
      • Take 1 µl of sample immediately, add with 2 µl nf H2O and 3 µl 2x RNA LD
      • Store at -80 °C
      • What still needs to be done:
        • Cook samples at 95 °C for 10 minutes and put directly on ice afterwards (to prevent refolding)
        • Load on Gel and quantify
Tuesday 6th
  • Inoculation of Rosetta pSB1C3-His-MBP-Lbu-Cas13a-WT and Rosetta p2CT-His-MBP-Lsh-Cas13a-WT overnight cultures in each 1 l LBCarb + Cm medium
  • Protocol:
  • Participants: Sven, Patrick
  • Preparation of glycerol stocks for DH5α pSB1C3-BBa_K1319004 cPCR and DH5α pSB1C3-BBa_K1319008
  • Protocol: Glycerol cryo stocks
  • Participants: Sven, Patrick
  • Minipreparation of DH5α pSB-Cas13a-Lwa cPCR colony 5, DH5α pSB1C3-BBa_K1319004 and DH5α pSB1C3-BBa_K1319008
  • Protocol: Miniprep
  • Participants: Sven, Patrick
Wednesday 7th
  • Harvesting of bacteria with overexpressed Cas13a Lbu and Cas13a Lsh
  • Protocol: Äkta purification
  • Participants: Ludwig
  • Preparation of 5 ml LBCm overnight culture of DH5α pSB-Cas13a-Lwa cPCR colony 5
  • Protocol: -
  • Participants: Jorge
Thursday 8th
  • Preparation of wash and elution buffer for Cas13a protein purification
  • Protocol: Äkta protein purification
  • Participants: Ludwig, Erika
  • Observations:
    • pH was checked and adapted after adding of TCEP
  • Purification of Cas13a Lbu and Cas13a Lsh (Affinity chromatography)
  • Protocol: Äkta protein purification
  • Participants: Ludwig, Erika, Sven

  • Results

    xxxChange
Friday 9th
  • PCR of pSB1C3-BBa_K1319008 (TEV) for adding a His tag
  • Protocol: PCR
  • Participants: Chris
  • Observations:
    • Primers: p-TEV-His-fwd and p-TEV-His-rev
    • Procedure was follwed by PCR cleanup and DpnI digestion

  • Results

    xxxChange
Monday 12th
  • Purification of Cas13a Lbu and Cas13a Lsh (Size exclusion chromatography)
  • Protocol: Äkta purification
  • Participants: Sven, Ludwig, Benedikt, Milica
  • Observations:
    • Cas13a Lbu and Cas13a were upconcentrated to 1 ml each (centrifugal filters: MWCO: 30 kDa). 2 runs for each protein were necessary

  • Results

    xxxChange
  • PCR of Lwa part2b (reordered gBlock without internal BsaI restriction site)
  • Protocol:
  • Participants: Sven, Ludwig, Benedikt, Milica
  • Observations:
    • Primers: VF, VR
    • Procedure was follwed by PCR cleanup

  • Results

    xxxChange
  • Assembly of Lwa part1, part2b and pSB1C3-ccdB with GoldenGate and transformation in DH5α
  • Protocol: GoldenGate Cloning
  • Participants: Sven, Ludwig, Benedikt, Milica
  • Ligation of pSB1C3-BBa_K1319008 (TEV) after adding the His tag by overhang PCR
  • Protocol:
  • Participants: Sven, Ludwig, Benedikt, Milica
  • Observations:
    • Procedure was follwed by chemicial transformation in DH5α and plating on LbCm agar plates
Tuesday 13th
  • Colony PCR of DH5α pSB-Cas13a-Lwa from GoldenGate assembly (with part2b)
  • Protocol:
  • Participants: Ludwig, Chris

  • Results

  • cPCR showed positive results. Cloning seemed to work, although there was the internal BsaI restriction site
    • xxxChange
  • Redone: Transformation of 15 µl ligation sample (of pSB1C3-BBa_K1319008 (TEV) after adding the His tag by overhang PCR)
  • Protocol:
  • Participants: Ludwig, Chris
  • Observations:
    • Colonies were visible the next day this time. 2 colonies were picked and resuspended in each 5 ml LBCm for overnight cultures
  • Preparation of 5 ml LBCm overnight culture of DH5α pSB-Cas13a-Lwa cPCR colony 5
  • Protocol:
  • Participants:
  • Observations:
    • Inoculation from cyro stock

  • Results

  • Lysis-tests with acidic phenol chlorophorm extraction
  • Protocol: Alkaline lysis Protocol: Phenol-Chlorophorm extraction Protocol: Trizol Reagenz protocol
  • Participants: Julian, Patrick
  • Notes:
    • Three lysis methods were used: Alkaline Lysis, heat lysis with SDS and Trizol-Lyse.
    • No vortexing for lysis (may break up cells and won't be available in our device), only room-temperature (apart form SDS as reference)
    • Cells had OD of 5.97 -> 4.78*10^9 cells according to http://www.genomics.agilent.com/biocalculators/calcODBacterial.jsp?_requestid=1029158
    • Cells had OD of 5.97 -> 4.78*10^9 cells according to http://www.genomics.agilent.com/biocalculators/calcODBacterial.jsp?_requestid=1029158
      -> dilluted 1:100 (to V=10mL) and pelleted 209 miL to get to 10^7 cells
    • A
    B
    C
    D
    1
    		MethodSDSNaOHTRIzol
    2
    		prepare1 mL TES (10mM Tris-HCl, 10mM EDTA, pH 8.0, 2% SDS -> TE-Buffer + SDS...
    3
    		----Lysis1x3x1x
    4
    		separate to tubes Spin 300xG, 5 min ->Ice, TROzol to -20! (100 miL at OD 1 -> 10^7 cells, Max. according to TRIzolPaper100 miL bactSusp100 miL bact-susp100 miL bact-suspension
    5
    		No washing step, bad for mRNA (TRIzolP.) 300 µl (0.45 ml) TES resuspend pellet
    6
    		12,5 uL Inhibitor (40k U/ml, 1 U/miL required)15 min, 75 °C lysis, vortexone probe after another
    7
    		(Addition of inhibitor); Transform total volume to gel tuberesuspend thorougly in 66 µl (100 miL) Solution1
    8
    		132 µl (200 miL) Sol2, INVERT 4x
    9
    		wait 0, 1 ,3 min
    10
    		add 99 µl (150 miL) Sol3, INVERT 4x
    11
    		Total volume: 297 µl (0.45 ml)
    12
    		--phenChloExtr
    13
    		measure pH, add acid...meassure Trizol PH, add acid if needed (pH <5)
    14
    		add 600 µl [same V (0.45 ml)] Phe:chloro.add 1 mL TRIzol to 10^7 cells
    15
    		5 min incubation
    16
    		add 0.2 mL CHLOROPHORM
    17
    		incub 2.5 (2-3) min, RT
    18
    		centri 12k g 15 min 4°C
    19
    		angle 45°, extract top phase carefully-> discard everything else, prot & DNA-> which Vol for the TRIzol aqueous phase?
    20
    		add 0.4 (0.375) ml Isopropanol, wait 10 min, RT
    21
    		centrifuge 10 min 12k , 4°C ->
    22
    		Aditional step, because RNA pellet was not entierly spinned down: Centrifugation (16000 rcf, 5 min, 4 °C)big-pellet assay in fridge, but 1. interphase was punctured/gel(DNA?) stuck to pipette 2. s.u.
    23
    		Aditional step, because suspected RNA fog was not entirely spinned down: remove upper and lower RNA fog phase
    24
    		Additional step: Centrifugation (16000 rcf, 5 min, 4 °C)
    25
    		No RNA pellet or RNA fog any more :(
    26
    		Addition of 400 µl isopropanol -> No RNA pellet or fog any more ->prob. removed too much
    27
    		discard supernatant -> RNA in gel-like pellet!-> gel hardly visible, nearly draged it out of tube (stuck to pipette...)
    28
    		resuspend in 0.75 ml 70 % (75% ideally) ethanol-> storeed RNA at -20°C is stable up to 1 year...
    29
    		vortex, centri 5 min 7,5k 4°
    30
    		discard supernatant, air dry for 5-10 min (do NOT let completely dry out)
    31
    		resuspend in RNAse free water 30 µl (20-50 miL) ->measurement...

  • Results

    No pellets seen during procedure, concentration too low for photometer -> repeat with more cells
Wednesday 14th
  • Up-concentration of Cas13a Lbu and Cas13 Lsh after SEC and concentration measurement
  • Protocol:
  • Participants: Chris, Patrick
  • Observations:
    • Centrifual filters: MWCO: 30 kDa
    • Concentrations were measured using a Bradford assay

  • Results

  • Cas13 Lbu: 746 mg/ml, Cas13 Lsh: 120 mg/ml
  • Miniprep of overnight culture of DH5α pSB-Cas13a-Lwa cPCR colony 5
  • Protocol: Minipreparation
  • Participants: Chris, Patrick
  • Observations:
    • Plasmid was again sent for sequencing with VR and VF

  • Results

  • First sequencing looked good
Thursday 15th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Friday 16th
  • Sending pSB-Cas13a-Lwa cPCR colony 5 for sequencing again with more primers
  • Protocol: Sequencing
  • Participants: Chris, Jorge
  • Observations:
    • Primer: p-seq-lwa 1-4
  • Preparation of first readout experiment with Cas13 Lbu and Cas13 Lsh
  • Protocol: Readout
  • Participants: Chris, Jorge
  • Observations:
    • Cleavage activity was tested with Aptamers (Malachite, Spinach) and RNase alert

  • Results

  • The Cas13a proteins were activated and cleaved the aptamers. This resulted in a decrease in fluorescence intensity.
    • xxxChange
  • Lysis-tests with acidic phenol chlorophorm extraction
  • Protocol: Lysis test pipetting scheme Protocol: Phenol-Chlorophorm extraction
  • Participants: Julian, Patrick
  • Notes:
    • Cell suspension had OD600 (extrapolated) of 2,220 -> 1.78 x 10^9 cells/ml according to http://www.genomics.agilent.com/biocalculators/calcODBacterial.jsp?_requestid=1029158
    • Experiments were done with 108 and 109 cells per sample, because it failed with 10^7 cells last time
    • Aspiration of supernatant in phenol-chlorophorm extraction: about 2/3 aspirated (200 µl) and 1/3 (100 µl) left to not destroy interphase
    • No pellet after RNA precipitation (-80 °C) and subsequent centrifugation in:
      • NaOH, 1 min, 10^8
      • SDS, 10^9
      • Trizol, 10^8
    • Mistakes:
      • SDS, 10^9: aspirated 100 µl instead of 200 µl supernatant
      • NaOH, 10^8, 3 min: contamination with DNA (<10% DNA interphase transfered)
      • NaOH, 10^9, 3 min: added + 200 µl of SDS 109 tube
    • ! Did not calibrate alkaline lysis solution 3 to given pH -> probably more RNA degradation
    • ! Did store RNA at -20 °C instead of -80 -> probably more RNA degradation

  • Results

    Sample
    [] [µg/ml]
    adjusted C**
    A260/A280
    A260/A230
    A230 [A]
    A260 [A]
    A280 [A]
    A320 [A]
    1
    		SDS, 10^8 cells36.8551.7042.0440.0470.0940.0560.002
    2
    		SDS, 10^9 cells*1193571.6932.0690.1460.3000.1780.002
    3
    		NaOH, 0 min, 10^8 cells35.2531.7252.1460.0420.0890.0520.001
    4
    		NaOH, 0 min, 10^9 cells5868791.9231.9930.7401.4700.7670.005
    5
    		NaOH, 1 min, 10^8 cells73.61101.2350.5860.3170.1870.1520.003
    6
    		NaOH, 1 min, 10^9 cells3705551.9451.9580.4750.9270.4780.003
    7
    		NaOH, 3 min, 10^8 cells*2593891.8491.4910.4390.6520.3550.005
    8
    		NaOH, 3 min, 10^9 cells*2754131.8502.2120.3160.6930.3680.005
    9
    		TRIzol, 10^8 cells1942651.3620.2212.2000.4870.3580.002
    10
    		TRIzol, 10^9 cells1602181.4290.4880.8220.4030.2830.003
  • Measured [RNA] by Implen Nanophotometer (20170616).
  • **adjusted for different supernatant V aspirated at extraction step
  • Barplots of measure RNA concentrations
    • see also "Denaturing PAGE for ssRNA" Monday, the 26th
Monday 19th
  • Preparation of 5 ml LBCm overnight culture of DH5α pSB-Cas13a-Lwa cPCR colony 5
  • Protocol: Overnight culture
  • Participants: Ludwig
  • Observations:
    • Inoculation from cryo stock
  • Preparation of 5 ml LBCm overnight culture of DH5α pSB1C3-BBa_K1319008-His (TEV) 1 and 2
  • Protocol: Overnight culture
  • Participants: Ludwig
  • Observations:
    • Inoculation from cryo stock
  • Electro transformation of pSB1C3-Cas13a-Lwa (cPCR colony 5) into E. coli BL21star
  • Protocol: Electro transformation
  • Participants: Ludwig, Chris
  • Observations:
    • pSB1C3-Cas13a-Lwa (cPCR colony 5) was verified by sequencing and is named now pSB1C3-Cas13a-Lwa
Tuesday 20th
  • Minipreparation of DH5α pSB-Cas13a-Lwa cPCR colony 5
  • Protocol: Minipreparation
  • Participants: Ludwig, Chris
  • Observations:
    • The plasmid was sent for sequencing with VR, VF, p-seq-lwa 1-4
  • Minipreparation of DH5α pSB1C3-BBa_K1319008-His (TEV) 1 and 2
  • Protocol: Minipreparation
  • Participants: Ludwig, Chris
  • Observations:
    • The plasmids were sent for sequencing with VR, VF
  • Preparation of 5 ml LBCm overnight culture of one picked BL21star pSB1C3-Cas13a-Lwa colony
  • Protocol: Overnight culture
  • Participants: Ludwig, Chris
Wednesday 21st
  • Minipreparation of BL21star pSB1C3-Cas13a-Lwa overnight culture and send for sequencing
  • Protocol: Minipreparation
  • Participants: Erika, Igor, Milica
  • Observations:
    • Primers: VF, VR, p-seq-lwa 1-4
  • Preparation of glycerol stock for BL21star pSB1C3-Cas13a-Lwa
  • Protocol: Glycerol stock
  • Participants: Erika, Igor, Milica
Thursday 22nd
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Friday 23rd
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
  • Time-series of alkaline lysis, in triplets
  • Protocol: Lysis test pipetting scheme
  • Participants: Julian, Patrick
  • Notes:
    • Did alkaline Lysis with inbubation/lysis times of 0 min (~10 sec ), 1 min, 3 min, 10 min, Heat+SDS as reference, no Trizol
    • Cell suspension with OD600 (extrapolated) of 2.182 -> 1.78 x 109 cells/ml
    • Phenol-Chlorophorm Precipitation at -80 was done until Wednesday

  • Results

    see "Denaturing PAGE for ssRNA" Thursday (not Monday!), the 29th
Monday 26th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
  • Gel analysis of extracted RNA (July 16th !)
  • Protocol: Denaturing PAGE for ssRNA
  • Participants: Julian, Patrick
  • Notes:
    • see Friday, 16th

  • Results

    Gel with degraded RNA
Tuesday 27th
  • Electro Transformation of pSB1C3-BBa_K1319008-His (TEV) in BL21star and plating on LBCm agar plates
  • Protocol: Electro transformation
  • Participants: Ludwig, Chris
  • Observations:
    • 1 µl was used
  • Preparation of 5 ml LBCm + Carb overnight culture of E. coli Rosetta p2CT-His-MBP-Lbu-Cas13a-WT
  • Protocol: Overnight culture
  • Participants: Ludwig, Chris
  • Preparation of 5 ml LBCm overnight culture of E. coli DH5α pSB1C3-BBa_K1319008-His 2
  • Protocol: Overnight culture
  • Participants: Ludwig, Chris
Wednesday 28th
  • Inoculation of Rosetta p2CT-His-MBP-Lbu-Cas13a-WT in 4x 1 l 2xYTCm + Carb medium for expression of Cas13a Lbu
  • Protocol: Äkta protein purification
  • Participants: Chris, Ludwig,Jorge
  • Minipreparation of DH5α pSB1C3-BBa_K1319008-His 2 and send for sequencing
  • Protocol: Minipreparation and Sequencing
  • Participants: Chris, Ludwig, Jorge
  • Observations:
    • Primers: VF, VR
  • Preparation of 5 ml LBCm overnight culture of BL21star pSB1C3-BBa_K1319008-His from picked colony
  • Protocol: Overnight culture
  • Participants: Chris, Ludwig, Jorge
  • Plating of DH5α pSB1C3-Cas13a-Lwa glycerol stock on LBCm agar plates
  • Protocol:
  • Participants:
  • Observations:
    • Colonies should be checked if they grow the same and a cPCR should confirm that there is only one plasmid
Thursday 29th
  • Preparation of glycerol stock of BL21star pSB1C3-BBa_K1319008-His
  • Protocol: Glycerol stock
  • Participants: Ludwig, Chris
  • Observations:
    • From this glycerol stock a 5 ml LBCm overnight culture was prepared
    • A 5 ml LBCm overnight culture was prepared directly from this cryo stock
  • Harvesting of Rosetta p2CT-His-MBP-Lbu-Cas13a-WT with overexpressed Cas13a Lbu
  • Protocol: Äkta protein purification
  • Participants: Ludwig, Chris
  • Observations:
    • The pellet was stored at -80 °C
  • Colony PCR of DH5α pSB1C3-Cas13a-Lwa (plated glycerol stock)
  • Protocol: cPCR
  • Participants: Ludwig, Chris

  • Results

    xxxChange
  • Time-series of alkaline lysis, in triplets (continued)
  • Protocol: Phenol-Chlorophorm extraction
  • Participants: Patrick
  • Notes:
    • Continuing after -80 °C precipitation
    • Extremely big pellets after 99 % EtOH precipitation for all samples with SDS: Possibly contaminated with DNA

  • Results

    • SDS 10⁹ seems to have lower yield than NaOH -> maybe more SDS? (10⁸ has no problems...)
    • NaOH is done after "0" minutes (about 10 sec for adding base to all eppis and turning 3 times)
    • Sample
      Number of sample
      [] [µg/ml]
      A260/A280
      A260/A230
      A230 [A]
      A260 [A]
      A280 [A]
      A320 [A]
      1
      		SDS, 10^8 cells162.42.0800.1181.3430.1740.0930.018
      2
      		256.82.0290.1281.1230.1530.0810.011
      3
      		347.61.9830.1021.1780.1270.0680.008
      4
      		SDS, 10^9 cells13541.782>0.4>2.50.9130.5250.029
      5
      		22601.8570.2952.2220.6650.3650.015
      6
      		33761.8010.3922.4220.9620.5440.022
      7
      		NaOH, 0 min, 10^8 cells139.21.6902.0420.0560.1060.0660.008
      8
      		253.61.6141.8870.0860.1490.0980.015
      9
      		368.81.7202.4570.0700.1720.1000.000
      10
      		NaOH, 0 min, 10^9 cells16291.9112.4660.6431.5780.8280.005
      11
      		217.61.6922.2000.0190.0430.025-0.001
      12
      		36191.8972.4530.6361.5530.8210.005
      13
      		NaOH, 1 min, 10^8 cells1521.8062.2030.0590.1300.0720.000
      14
      		233.21.7292.3710.0350.0830.0480.000
      15
      		358.41.7382.3930.0610.1460.0840.000
      16
      		NaOH, 1 min, 10^9 cells16061.8932.4690.6201.5220.8070.006
      17
      		26271.8982.4310.6511.5740.8320.006
      18
      		322.41.8062.4350.0230.0560.0310.000
      19
      		NaOH, 3 min, 10^8 cells1441.8642.4440.0440.1090.058-0.001
      20
      		261.21.7792.5930.0580.1520.085-0.001
      21
      		35.61.4002.8000.0040.0130.009-0.001
      22
      		NaOH, 3 min, 10^9 cells16191.9082.4320.6431.5540.8180.007
      23
      		25701.9272.4680.5831.4300.7450.006
      24
      		320.41.7592.4290.0200.0500.028-0.001
      25
      		NaOH, 10 min, 10^8 cells110.81.5882.2500.0120.0270.0170.000
      26
      		28.81.5713.1430.0060.0210.013-0.001
      27
      		34.41.3753.6670.0020.0100.007-0.001
      28
      		NaOH, 10 min, 10^9 cells15031.9162.4990.5081.2620.6610.005
      29
      		25641.9112.4870.5721.4150.7430.005
      30
      		36111.9022.4670.6251.5330.8090.006
Friday 30th
  • Minipreparation of BL21star pSB1C3-BBa_K1319008-His
  • Protocol: Minipreparation
  • Participants: Max
  • Observations:
    • The plasmid was sent for sequencing with primers VR and VF
  • Gel analysis of alkaline-extracted RNA time-series (July 23th !)
  • Protocol: Denaturing PAGE for ssRNA
  • Participants: Julian, Patrick
  • Notes:
    • see Friday, 23th

  • Results

    Gel RNA degrading with lysis time
Monday 3rd
  • Preparation of 5 ml LBCm overnight culture of BL21star pSB-Cas13a-Lwa
  • Protocol: Overnight culture
  • Participants: Max
Tuesday 4th
  • Purification of Cas13a Lbu
  • Protocol: Ni NTA agarose purification
  • Participants: Max, Dawa
  • Observations:
    • Fresh DTT was added to the lysis buffer
    • 1 ml Ni-NTA agarose was used per 4 ml lysate
    • Sample was incubated 1 h at 4 °C while shaking
    • The procedure was followed by TEV cleavage overnigth while dialyzing

  • Results

    xxxChange
Wednesday 5th
  • Harvesting of BL21star pSB-Cas13a-Lwa with overexpressed Cas13a Lwa
  • Protocol: Ni NTA protein purification
  • Participants: Max, Dawa, Erika
  • Observations:
    • Pellet was incubated for 1 h at -80 °C before it was used for purification

  • Results

  • Purification of Cas13a Lwa
  • Protocol: Ni NTA agarose purification
  • Participants: Max, Dawa, Erika
  • Observations:
    • Fresh DTT was added to the lysis buffer
    • 1 ml Ni-NTA agarose was used per 4 ml lysate
    • Sample was incubated 1 h at 4 °C while shaking

  • Results

    xxxChange
  • Precipitation of TEV plasmid for European meetup in Delft
  • Protocol: -
  • Participants: Max, Dawa, Erika
  • Observations:
    • pSB1C3-BBa_K1319008-His was lyophylized by Ethanol precipitation and evaporation
Thursday 6th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Friday 7th
  • Reverse Ni-NTA purification of cut Cas13a Lbu
  • Protocol: Ni NTA agarose protein purification
  • Participants: Rob
  • Observations:
    • Ni NTA agarose binds the cleaved His-MBP tag, Cas13a Lbu is in the flow-through

  • Results

    xxxChange
Monday 10th
  • Preparation of 50 ml LBCm overnight culture of BL21star pSB1C3-BBa_K1319008-His (TEV)
  • Protocol: Overnight culture
  • Participants: Rob, Ludwig, Rob
Tuesday 11th
  • Inoculation of BL21star pSB1C3-BBa_K1319008-His (TEV) in 3x 1 l 2xYTCm medium for expression of the TEV protease
  • Protocol: Äkta protein purification
  • Participants: Ludwig, Max
  • Observations:
    • Induction with 1 mM IPTG at OD600 of 0.6
    • Expression at 16 °C overnight
  • RNase alert assay with Cas13a Lbu and Cas13a Lsh
  • Protocol: Cas13a activity assay
  • Participants: Igor, Rob

  • Results

    xxxChange
Wednesday 12th
  • Harvesting of BL21star pSB1C3-BBa_K1319008-His with overexpressed TEV protease
  • Protocol: Äkta protein purification
  • Participants: Ludwig, Milica
Thursday 13th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Friday 14th
  • Cas13a activity assay with RNase alert
  • Protocol: Cas13 activity assay
  • Participants: Dawa, Rob

  • Results

    xxxChange
Monday 17th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Tuesday 18th
  • Preparation of 40 ml LBCm + Carb overnight culture of Rosetta p2CT-His-MBP-Lbu-Cas13a-WT
  • Protocol: Äkta protein purification
  • Participants: Chris, Max, Dawa
Wednesday 19th
  • Inoculation of Rosetta p2CT-His-MBP-Lbu-Cas13a-WT in 3x 1 l 2xYTCm + Carb medium for expression of Cas13a Lbu
  • Protocol: Äkta protein purification
  • Participants: Max, Dawa
Thursday 20th
  • Size exclusion chromatography of affinity purified TEV protease
  • Protocol: Äkta protein purification
  • Participants: Max, Sven, Ludwig

  • Results

    xxxChange
  • Harvesting of Rosetta p2CT-His-MBP-Lbu-Cas13a-WT with overexpressed Cas13 Lbu
  • Protocol: Äkta protein purification
  • Participants: Max, Sven, Ludwig
  • In vitro Transcription of target RNA, Lbu crRNA and Lsh crRNA
  • Protocol: In vitro Transcription
  • Participants: Igor
  • Observations:
    • Procedure was followed by DNAase treatment and phenol chloroform extraction

  • Results

    xxxChange
Friday 21st
  • PCR of gBlock of aeBlue
  • Protocol: PCR
  • Participants: Chris
Monday 24th
  • Digestion of insert and pSB1C3 with Pst1-HF and EcoRI-HF
  • Protocol: Digestion and Ligation
  • Participants: Chris
  • Observations:
    • Plasmid was dephosporylated with arctic phosphatase and insert was ligated with plasmid overnight at 16 °C
Tuesday 25th
  • Electro transformation of aeBlue ligation sample in E. coli Turbo cells
  • Protocol: Electro transformation
  • Participants: Max
Wednesday 26th
  • Colony PCR of plated pSB1C3-aeBlue
  • Protocol: cPCR
  • Participants: Max

  • Results

    xxxChange
  • Preparation of 5 ml LBCm overnight cultures (picked colonies of aeBlue ligation)
  • Protocol: Overnight culture
  • Participants: Max
Thursday 27th
Friday 28th
  • Upconcentration of TEV protease to 2 mg/ml and stored in TEV storage buffer
  • Protocol: Äkta protein purification
  • Participants: Max
Monday 31th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Tuesday 1st
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Wednesday 2nd
  • InterLab Study calibration
  • Protocol: IGEM HQ InterLab Study Protocol
  • Participants: Dawafuti Erika
  • Observations:
    • Patch length correction was on in the plate reader.
    • Calibration for the plate reader experiment done with LUDOX and Fluorescein.
    • We saw no difference between LUDOX and water when measuring at an OD of 600 nm.
    • For the fluorescein measurement, the best gain was 600 using 485 nm/520 nm.
    • Plasmids from the plate 7 (positive and negative control, and devices 1-6) were resuspended.
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Thursday 3rd
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Friday 4th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Monday 7th
  • InterLab Study Day 1: Transformation
  • Protocol: IGEM HQ InterLab Study Protocol
  • Participants: Dawafuti Erika
  • Observations:
    • Each plasmids was transformed two times: with chemical transformation and with electroporation, for a total of 16 plasmids.

  • Results

    • Every plate except the ones for Device 1 had colonies.
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Tuesday 8th
  • InterLab Study Day 2: Colony transfer to liquid culture
  • Protocol: IGEM HQ InterLab Study Protocol
  • Participants: Dawafuti Erika
  • Observations:
    • Device 1 was again transformed, as no colonies grew on the plate.
    • Liquid cultures for the 7 plasmids whose plates showed colonies (all except Device 1) were made in duplicate.
    • The plates were stored at -4 °C.
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Wednesday 9th
  • InterLab Study Day 3: Plate reader experiments
  • Protocol: IGEM HQ InterLab Study Protocol
  • Participants: Dawafuti Erika
  • Observations:
    • After measuring the OD600 of the O/N cultures, they were diluted with LB+Cm to an OD600 of 0.02, in duplicates.
    • The absorbance and fluorescence of the diluted cultures were measured at 0, 2, 4 and 6 hours after the cultures were set.
    • Patch length correction was on in the plate reader.
    • Device 1 was again transformed, as no colonies grew on the plate.

  • Results

    • Transformation of Device 1 was successful.
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Thursday 10th
  • InterLab Study Day 4: Preparation of O/N cultures for plate reader experiments
  • Protocol: IGEM HQ InterLab Study Protocol
  • Participants: Dawafuti Erika
  • Observations:
    • O/N liquid cultures in duplicate were set up from all 8 Devices.
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Friday 11th
  • InterLab Study Day 5: Plate reader calibration and experiments
  • Protocol: IGEM HQ InterLab Study Protocol
  • Participants: Dawafuti Erika
  • Observations:
    • Patch length correction was off in the plate reader.
    • Calibration for the plate reader experiment done with LUDOX and Fluorescein.
    • We saw the difference in measurements between LUDOX and water.
    • After measuring the OD600 of the O/N cultures, they were diluted with LB+Cm to an OD600 of 0.02, in duplicates.
    • The absorbance and fluorescence of the diluted cultures were measured at 0, 2, 4 and 6 hours after the cultures were set.

  • Results

  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Monday 14th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Tuesday 15th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Wednesday 16th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Thursday 17th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Friday 18th
Sunday 20th
Monday 21st
  • FINA extraction for DNA with E. coli W3110 pre-purified gDNA
  • Protocol: FINA Extraction for DNA
  • Participants: Jorge
  • Observations:
    • The gDNA was pre-purified with the Phenol/Chloroform method.
    • The gDNA solution was diluted 1:100, 1:1000, 1:100000, 1:10e6 and 1:10e8. Each dilution was then used as a sample for the FINA extraction.
    • From each dilution, two extractions were performed. One as in protocol (1N-5N), the other without the NaOH washing step (1-5).
    • As negative control (KK) FINA extraction was performed as in protocol with nuclease free water as sample.
  • Q5-PCR for beta-galactosidase from E. coli with FINA-DNA samples and purified gDNA from 21.08.17
  • Protocol: PCR amplification with Q5 Master Mix
  • Participants: Jorge
  • Observations
    • For the membranes, no template was directly pipette. Instead, the membranes were put in the tube.
    • p-bGal_N_N was used as forward primer
    • p-bGal_N_C was used as reverse primer
    • Annealing was performed at 61 ºC
    • The amplification step was 90 s long
    • A 1.5% agarose gel was run in TAE-buffer at 110 V for 80 min to check for PCR-products
    • Quick-Load Purple 2-Log DNA Ladder (N0550S) was used as ladder

  • Results

    • 1N-5N: 1:100-1:10e8 dilutions FINA extraction performed as in protocol.
    • 1-5: 1:100-1:10e8 dilutions FINA extraction without washing step.
    • 1K-5K: 1:100-1:10e8 dilutions.
    • KK: negative control for FINA extraction (nuclease free water as sample).
Tuesday 22nd
  • FINA extraction for DNA with E. coli W3110 cell culture
  • Protocol: FINA Extraction for DNA
  • Participants: Jorge
  • Observations:
    • The extraction was done when the culture had an OD600 of 1.2.
    • The cells were diluted 1:1, 1:100 and 1:1000 before the extraction.
    • Each dilution was used as sample twice: as in protocol (1N-3N and P1N-P3N) and washed with 300 ul lysis buffer from RNA extraction kit (Norgen, P/N 17200) (1G-3G and P1G-P3G).
    • Membranes 1N-3N and 1G-3G were put in 40 ul nuclease free water and incubated at RT for 10 min in order for the bounded DNA to go into solution.
  • Q5-PCR for beta-galactosidase from E. coli with FINA-DNA samples from 22.08.17
  • Protocol: PCR amplification with Q5 Master Mix
  • Participants: Jorge
  • Observations
    • The membranes from the FINA extraction were directly put into the PCR-mix. No template DNA was directly pipetted.
    • p-bGal_N_N was used as forward primer.
    • p-bGal_N_C was used as reverse primer.
    • Annealing was performed at 61 ºC.
    • The amplification step was 90 s long.
    • A 1.5% agarose gel was run in TAE-buffer at 110 V for 80 min to check for PCR-products.
    • Quick-Load Purple 2-Log DNA Ladder (N0550S) was used as ladder

  • Results

    • 1N-3N: 1:1-1:1000 dilutions FINA extraction performed as in protocol, loaded in gel after elution (no PCR).
    • 1G-3G: 1:1-1:1000 dilutions FINA extraction performed with lysis buffer as washing step, loaded in gel after elution (no PCR).
    • P1N-P3N: 1:1-1:1000 dilutions FINA extraction performed as in protocol.
    • P1G-P3G: 1:1-1:1000 dilutions FINA extraction with lysis buffer as washing buffer.
    • KK: negative control for FINA extraction (nuclease free water as sample).
Wednesday 23rd
  • FINA extraction for DNA with E. coli W3110 cell culture
  • Protocol: FINA Extraction for DNA
  • Participants: Jorge, Benedikt
  • Observations:
    • The extraction was done when the culture had an OD600 of 2.33.
    • The cells were diluted 1:1, 1:100, 1:10000, 1:10e6 and 1:10e8 before the extraction.
    • Each dilution was used as a sample for the FINA extraction. In addition, three 1:1 dilutions were used as samples for FINA extraction: two according to protocol and the third with 300 ul lysis buffer as washing buffer. One of the 1:1 membranes was put in nuclease free water and incubated at RT for 20 min in order for the bounded DNA to go into solution.
  • Q5-PCR for beta-galactosidase from E. coli with FINA-DNA samples from 23.08.17
  • Protocol: PCR amplification with Q5 Master Mix
  • Participants: Jorge, Benedikt
  • Observations
    • The membranes from the FINA extraction were directly put into the PCR-mix. No template DNA was directly pipetted.
    • p-bGal_N_N was used as forward primer.
    • p-bGal_N_C was used as reverse primer.
    • Annealing was performed at 61 ºC.
    • The amplification step was 90 s long.
    • A 1.5% agarose gel was run in TAE-buffer at 110 V for 80 min to check for PCR-products.
    • Afte the PCR, only 2,5,L and KK were loaded in the gel, as the other tubes had no liquid.
    • The gel was loaded before it was submerged in TAE-buffer.
    • Quick-Load Purple 2-Log DNA Ladder (N0550S) was used as ladder

  • Results

    • 2: 1:100 dilution FINA extraction performed as in protocol
    • L: 1:1 dilution FINA extraction performed with 300 ul lysis buffer as washing buffer.
    • KK: negative control for FINA extraction (LB-medium as sample).
Thursday 24th
  • FINA extraction for DNA with E. coli W3110 cell culture (Repetition of the experiment from 23.08.17)
  • Protocol: FINA extraction for DNA
  • Participants: Jorge, Benedikt
  • Observations:
    • As there were problems with the gel from the 23th, we repeated the experiment.
    • The extraction was done when the culture had an OD600 of 2.78.
    • The cells were diluted 1:1, 1:100, 1:10000, 1:10e6 and 1:10e8 before the extraction.
    • Each dilution was used as a sample for the FINA extraction. In addition, three 1:1 dilutions were used as samples for FINA extraction: two according to protocol and the third with 300 ul lysis buffer as washing buffer. One of the 1:1 membranes was put in nuclease free water and incubated at RT for 20 min in order for the bounded DNA to go into solution.
  • Q5-PCR for beta-galactosidase from E. coli with FINA-DNA samples from 24.08.17
  • Protocol: PCR amplification with Q5 Master Mix
  • Participants: Jorge, Benedikt
  • Observations
    • The membranes from the FINA extraction were directly put into the PCR-mix. No template DNA was directly pipetted.
    • p-bGal_N_N was used as forward primer.
    • p-bGal_N_C was used as reverse primer.
    • Annealing was performed at 61 ºC.
    • The amplification step was 90 s long.
    • A 1.5% agarose gel was run in TAE-buffer at 110 V for 80 min to check for PCR-products.
    • Quick-Load Purple 2-Log DNA Ladder (N0550S) was used as ladder

  • Results

    • 1-5: 1:1-1:10e8 dilutions FINA extraction performed as in protocol
    • L: 1:1 dilution FINA extraction performed with 300 ul lysis buffer as washing buffer.
    • KK: negative control for FINA extraction (LB-medium as sample).
  • Total RNA purification from E. coli Rosetta2 DE3 p2CT-His-MBP-Lsh_C13a_WT with Norgen kit
  • Protocol: Total RNA Purification with Norgen Kit
  • Participants: Jorge, Benedikt
  • Observations:
    • 2 tubes stored at -80 ºC. Labelled gRNA #1, gRNA #2.

  • Results

  • Phenol/Chloroform total RNA purification from E. coli Rosetta2 DE3 p2CT-His-MBP-Lsh_C13a_WT
  • Protocol: Phenol/Chloroform purification of RNA
  • Participants: Jorge
  • Observations:
    • The incubation step at -80 ºC was done, O/N.
    • 2 tubes stored at -80 ºC. Labelled gRNA PEC #1, gRNA PEC #2.

  • Results

Friday 25th
  • Continuation of Phenol/Chloroform total RNA purification from E. coli Rosetta2 DE3 p2CT-His-MBP-Lsh_C13a_WT from yesterday
  • Protocol: Phenol/Chloroform purification for RNA
  • Participants: Jorge
  • Observations:
    • See entry from 24.08.17
Monday 28th
Tuesday 29th
  • FINA extraction for DNA with E. coli Rosetta2 DE3 p2CT-His-MBP-Lsh_C13a_WT
  • Protocol: FINA extraction for DNA
  • Participants: Jorge
  • Observations:
    • The extraction was done when the culture had an OD600 of 0.88.
    • Na: Extraction as in protocol. Cl: Washed with 10 mM HCl instead of NaOH. H: incubated 10 min at 80 ºC. L: 10 min incubated in 1 mg/ml lysozyme. LH: 10 min incubated in 1 mg/ml lysozyme at 80 ºC.
  • Q5-PCR for beta-galactosidase from E. coli with FINA-DNA samples from 24.08.17
  • Protocol: PCR amplification with Q5 Master Mix
  • Participants: Jorge
  • Observations
    • The membranes from the FINA extraction were directly put into the PCR-mix. No template DNA was directly pipetted.
    • p-bGal_N_N was used as forward primer.
    • p-bGal_N_C was used as reverse primer.
    • Annealing was performed at 61 ºC.
    • The amplification step was 90 s long.
    • A 1.5% agarose gel was run in TAE-buffer at 110 V for 80 min to check for PCR-products.
    • Quick-Load Purple 2-Log DNA Ladder (N0550S) was used as ladder

  • Results

    See Q5-PCR from 30.08.17
  • FINA extraction for RNA with purified total RNA (gRNA PEC #1)
  • Protocol: FINA extraction for RNA
  • Participants: Jorge
  • Observations:
  • First strand cDNA synthesis from E. coli Rosetta2 DE3 p2CT-His-MBP-Lsh_C13a_WT purified total RNA (Phenol/Chloroform extraction from 24.08.17) and FINA-RNA eluate
  • Protocol: First strand cDNA synthesis
  • Participants: Jorge
  • Observations:
    • p-bGal_N_C was used as primer.
    • Sample gRNA PEC #1 from 24.08.17 was diluted 1:4 and used as sample P.
    • The eluate from the FINA extraction for RNA from 29.08.17 was used as sample S.
    • For both reactions (S and P), 6 ul sample were pipetted.
Wednesday 30th
  • Q5-PCR for beta-galactosidase from E. coli with cDNA samples from 29.08.17
  • Protocol: PCR amplification with Q5 Master Mix
  • Participants: Jorge
  • Observations:
    • p-bGal_N_N was used as forward primer.
    • p-bGal_N_C was used as reverse primer.
    • Annealing was performed at 61 ºC.
    • The amplification step was 90 s long.
    • A 1.5% agarose gel was run in TAE-buffer at 110 V for 80 min to check for PCR-products.
    • Quick-Load Purple 2-Log DNA Ladder (N0550S) was used as ladder

  • Results

    • Na: FINA extraction performed as in protocol.
    • Cl: FINA extraction performed with 10 mM HCl instead of NaOH.
    • L: Cell solution incubated 10 min in 1 mg/ml lysozyme. No washing step in FINA
    • H: Cell solution incubated 10 min at 80 ºC. No washing step in FINA
    • LH: Cell solution incubated 10 min at 80 ºC in 1 mg/ml lysozyme. No washing step in FINA
    • K-: negative control FINA (LB-medium used as sample).
    • P: 1:4 dilution of gRNA PEC #1 after reverse transcription.
    • S: 1:4 dilution of gRNA PEC #1 after FINA extraction for RNA and reverse transcription.
    • KK: Negative control for the cDNA synthesis (water instead of MuLV-RT used)
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Thursday 31st
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Friday 1st
  • Intein-Extein-Readout
  • Protocol: -
    • RNA extraction from induced Lbu-strain (continued)
    • Protocol: Denaturing PAGE for ssRNA
    • Participants: Dawa, Julian
    • Notes:
      • Induced pSB1C3_lbu with IPTG before lysis
      • Two cultures lysed with SDS + Heat only
      • No OD600 measurement

    • Results

      • A
        B
        C
        1
        		sampleconc ~Nanodrop260/280
        2
        		1 (non-induced)21921.8
        3
        		2 (non-induced)23201.8
        4
        		3 (induced)5081.69
        5
        		4 (induced)4421.66
        6
        		 1,2 showed precipitation of RNA ->measurement/gel wrong if not vortexed completely
      • A260/280 ration hints to impurities or DNA ->maybe due to too many cells
    Saturday 2nd
    Monday 4th
    • Total RNA extraction of E. coli Rosetta2 DE3 p2CT-His-MBP-Lsh_C13a_WT with Norgen kit
    • Protocol: Total RNA Purification with Norgen Kit
    • Participants: Jorge, Dawafuti
    • Observations:
      • Stored at -80 ºC as TRNA Kit #1-4
    • Phenol/Chloroform total RNA purification from E. coli Rosetta2 DE3 p2CT-His-MBP-Lsh_C13a_WT
    • Protocol: Protocol
    • Participants: Jorge, Dawafuti
    • Observations
      • Stored at -80 ºC as TRNA PC #1-4.

    • Results

      See results Urea-SDS-PAGE from 05.09.17.
    Tuesday 5th
    • Phenol/Chloroform total RNA purification from E. coli Rosetta2 DE3 p2CT-His-MBP-Lsh_C13a_WT (continued)
    • Protocol: Denaturing PAGE for ssRNA
    • Participants: Dawa, Jorge
    • Notes:
      • Ran urea gel

    • Results

      20170905_rna_1-8_Dawa_9_12_Jul.Tif
    • DNA Amplification Test using Recombinase Polymerase Amplification
    • Protocol: Recombinase Polymerase Amplification
    • Participants: Sven
    • Observations:
      • DNA Amplification with RPA was tested using the provided Control Reaction by TwistDx and our BioBrick His6-TEV plasmid incubation times of 40 minutes for Control and Sample 1 and 60 minutes for sample 2. Differences due to incubation time was tested.
      • RPA according to protocol
      • PCR cleanup according to protocol.

    • Results

      • Positive Control worked. RPA of TEV could not produce the whole (~1300 bp) construct but would produce amplicons of shorter size (100-500 bp) which was anticipated since TwistDx protocol says optimal amplicon length is 500 bp. Full length double-stranded DNA could consequently not be formed.

    Wednesday 6th
    • readout
    • Protocol: Protocol
    • Participants:
    • Observations
    • Results
    Thursday 7th
    • readout
    • Protocol: Protocol
    • Participants:
    • Observations
    • Results
    Friday 8th
    • readout
    • Protocol: Protocol
    • Participants:
    • Observations
    • Results
    • Test heat-only lysis (1,3 and 7 minutes))
    • Protocol: Phenol-Chlorophorm extraction
    • Participants: Julian
    • Notes:
      • Diluted E. coli culture to 10^8 cells/ml, used 300 uL as sample
      • Heat-only at 93 °C for 15 min, diluted wiht tap water to simulate actual usage scenario (saliva etc..)

    • Results

      Hardly any RNA detectable (concentration < 10 ug/ml). Abysmal 260/280-ratio
      -> Heat-only lysis is too inefficient for that low cell counts, use 10^9 next time.
    Monday 11th
    • readout
    • Protocol: Protocol
    • Participants:
    • Observations
    • Results
    Tuesday 12th
    • readout
    • Protocol: Protocol
    • Participants:
    • Observations
    • Results
    Wednesday 13th
    • readout
    • Protocol: Protocol
    • Participants:
    • Observations
    • Results
    • Heat-lysis with E. coli
    • Protocol: Phenol-Chlorophorm extraction
    • Participants: Julian
    • Notes:
      • Lysis at 90°C, used 300 uL of E. coli suspension diluted to 2*10^9 cells/ml
      • SDS-Lysis as reference was impossible with this cell count, interphase during extraction too broad

    • Results

      TODO table Extraction-yield is down 5- to 10- fold compared to SDS (calculated from SDS-lysis results with 10^9 suspension above)
    • RPA stability test and Reaction Temperature Screening on paper
    • Protocol: Recombinase Polymerase Amplification on Paper
    • Participants: Sven
    • Observations:
      • Preparation of stability assay of RPA on paper. Lyophilisation and storage.
      • RPA on paper according to protocol
    Friday 15th
    • Phenol/Chloroform total RNA purification from E. coli Rosetta2 DE3 p2CT-His-MBP-Lsh_C13a_WT
    • Protocol: Phenol/Chloroform purification of RNA
    • Participants: Jorge
    • Observations:
      • Two samples stored at -80 ºC labeled TRNA Lsh #1 15.09.17 and TRNA Lsh #2 15.09.17

    • Results

    • RPA stability test and Reaction Temperature Screening on paper
    • Protocol: Recombinase Polymerase Amplification on Paper
    • Participants: Sven
    • Observations:
      • Stability of RPA on paper was tested using His6-TEV plasmid after 0, 2 and 24 hours post-lyophilisation. Reaction temperature was tested at 37 °C as suggested by TwistDx and room temperature. Reaction was positively controlled using non-lyophilised reaction mixture.
      • RPA according to protocol
      • PCR cleanup according to protocol.

    • Results

      Lyophilisation of RPA on paper worked. Reaction seems to not be active at room temperature. Directly after lyophilisation, the reaction efficiency seems to be similar to fresh RPA reaction mixture. However, activity loss is observed already after 2 hours and after 24 hours, almost no activity is observable anymore.
      Gel Picture 20170915_Sven_RPA_time_stability
    Monday 18th
    • readout
    • Protocol: Protocol
    • Participants:
    • Observations
    • Results
    Tuesday 19th
    • readout
    • Protocol: Protocol
    • Participants:
    • Observations
    • Results
    • Heat-lysis with B. subtilis
    • Protocol: Phenol-Chlorophorm extraction
    • Participants: Julian
    • Notes:
        2x SDS-Heat lysis, 2x 90 °C (20 min)

    • Results

      SDS-lysis works a lot worse for gram+ bacteria, Heat-only now better/same (more incubation time...) Ration relatively bad, probably a lot of cell-wall impurities TODO table
    Wednesday 20th
    • readout
    • Protocol: Protocol
    • Participants:
    • Observations
    • Results
    Thurdsay 21st
    • readout
    • Protocol: Protocol
    • Participants:
    • Observations
    • Results
    Friday 22nd
    • readout
    • Protocol: Protocol
    • Participants:
    • Observations
    • Results
    Monday 25th
    • readout
    • Protocol: Protocol
    • Participants:
    • Observations
    • Results
    Tuesday 26th
    • readout
    • Protocol: Protocol
    • Participants:
    • Observations
    • Results
    Wednesday 27th
    • Phenol/Chloroform total RNA purification from E. coli Rosetta2 DE3 p2CT-His-MBP-Lsh_C13a_WT
    • Protocol: Phenol/Chloroform purification of RNA
    • Participants: Jorge
    • Observations:
      • Four samples stored at -80 ºC labeled TRNA Lsh #1 27.09.17 and TRNA Lsh #2 27.09.17, TRNA Lsh #3 27.09.17, TRNA Lsh #4 27.09.17.

    • Results

    • readout
    • Protocol: Protocol
    • Participants:
    • Observations
    • Results
    Thursday 28th
    • readout
    • Protocol: Protocol
    • Participants:
    • Observations
    • Results
    Friday 29th
    • FINA extraction for RNA with purified total RNA (TRNA Lsh #2-4 from 27.09.17)
    • Protocol: FINA Extraction for RNA
    • Participants: Jorge
    • Observations:
      • FINA was performed with 25 ul sample instead of 50 ul.
      • The extraction for TRNA #2 was performed as in protocol.
      • Before the FINA extraction, TRNA #3 was digested with DNase I according to DNase I Reaction Protocol
      • After the FINA extraction, the two eluate from TRNA #4 were digested with DNase I according to DNase I Reaction Protocol
    • First strand cDNA synthesis from E. coli Rosetta2 DE3 p2CT-His-MBP-Lsh_C13a_WT FINA-RNA eluate
    • Protocol: First strand cDNA synthesis
    • Participants: Jorge
    • Observations
      • pseq-Lsh-06rev was used as primer.
      • The eluates from the FINA extractions for RNA from 29.09.17 were used as samples.
      • From each pair of samples, one was used as a negative control (nuclease free water instead of MuLV-RT).
      • For all reactions, 6 ul sample were pipetted.
    • Q5-PCR for Lsh-fragment in E. coli Rosetta2 DE3 p2CT-His-MBP-Lsh_C13a_WT with cDNA samples from 29.09.17
    • Protocol: PCR amplification with Q5 Master Mix
    • Participants: Jorge
    • Observations:
      • pseq-Lsh-05fwd was used as forward primer.
      • pseq-Lsh-06rev was used as reverse primer.
      • Annealing was performed at 58 ºC.
      • The amplification step was 110 s long.
      • A 1.5% agarose gel was run in TAE-buffer at 110 V for 80 min to check for PCR-products.
      • Quick-Load Purple 2-Log DNA Ladder (N0550S) was used as ladder

    • Results

      • N: FINA extraction performed as in protocol.
      • NK: FINA extraction as in protocol. No RT in cDNA synthesis (negative control).
      • P: Digestion of DNA before FINA extraction.
      • PK: Digestion of DNA before FINA extraction. No RT in cDNA synthesis (negative control).
      • A: Digestion of DNA after FINA extraction.
      • AK: Digestion of DNA after FINA extraction. No RT in cDNA synthesis (negative control).
Monday 2nd
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Tuesday 3rd
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Wednesday 4th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Thursday 5th
Friday 6th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Monday 9th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Wednesday 11th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Thursday 12th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Friday 13th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Monday 16th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Tuesday 17th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Wednesday 18th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Thursday 19th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Friday 20th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Monday 23rd
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Tuesday 24th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Wednesday 25th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Thursday 26th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Friday 27th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Monday 30th
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results
Tuesday 31st
  • readout
  • Protocol: Protocol
  • Participants:
  • Observations
  • Results