Difference between revisions of "Team:Munich/LabJournal"
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<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <img src="AGAROSEGEL"> | + | <img src="AGAROSEGEL"> |
</li> | </li> | ||
</li> | </li> | ||
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<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <img src="AGAROSEGEL"> | + | <img src="AGAROSEGEL"> |
</li> | </li> | ||
</li> | </li> | ||
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<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <img src="GELIMAGE"> | + | <img src="GELIMAGE"> |
</li> | </li> | ||
</ul> | </ul> | ||
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<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <img src="PCR image (see 8.6.?)"> | + | <img src="PCR image (see 8.6.?)"> |
</li> | </li> | ||
</ul> | </ul> | ||
| Line 1,706: | Line 1,706: | ||
The Cas13a proteins were activated and cleaved the aptamers. This resulted in a decrease in fluorescence intensity. | The Cas13a proteins were activated and cleaved the aptamers. This resulted in a decrease in fluorescence intensity. | ||
</li> | </li> | ||
| − | <img src="A lot of images!!!!"> | + | <img src="A lot of images!!!!"> |
</li> | </li> | ||
</ul> | </ul> | ||
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<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <img src="GELIMAGEs.5.7."> | + | <img src="GELIMAGEs.5.7."> |
</li> | </li> | ||
</ul> | </ul> | ||
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<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <img src="GELIMAGE"> | + | <img src="GELIMAGE"> |
</li> | </li> | ||
</ul> | </ul> | ||
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<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <img src="graph"> | + | <img src="graph"> |
</li> | </li> | ||
</ul> | </ul> | ||
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<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <img src="GRAPH"> | + | <img src="GRAPH"> |
</li> | </li> | ||
| Line 2,753: | Line 2,753: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <img src="GELIMAGE maybe in next days"> | + | <img src="GELIMAGE maybe in next days"> |
</li> | </li> | ||
| Line 2,828: | Line 2,828: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <img src="GELIMAGE"> | + | <img src="GELIMAGE"> |
</li> | </li> | ||
| Line 2,881: | Line 2,881: | ||
<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"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">DNA Gel-Purification and Genomic DNA Purification</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">DNA Gel-Purification and Genomic DNA Purification</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 2,898: | Line 2,898: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Purification of b-Gal fragments from 1 % Agarose Gel <!-- | + | Purification of b-Gal fragments from 1 % Agarose Gel <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 2,908: | Line 2,908: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
DNA was lost during the purification | DNA was lost during the purification | ||
</li> | </li> | ||
</ul> | </ul> | ||
<div class="date">Wednesday 2nd</div> | <div class="date">Wednesday 2nd</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Q5-PCR and PCR-Cleanup kit</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Q5-PCR and PCR-Cleanup kit</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 2,930: | Line 2,930: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Repeated PCR of beta-Gal N and C Terminal fragments from 31.7. <!-- | + | Repeated PCR of beta-Gal N and C Terminal fragments from 31.7. <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 2,988: | Line 2,988: | ||
</ul> | </ul> | ||
<div class="date">Thursday 3rd</div> | <div class="date">Thursday 3rd</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Agarose-Gel</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Agarose-Gel</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 3,005: | Line 3,005: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Gel of purified PCR <!-- | + | Gel of purified PCR <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 3,015: | Line 3,015: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
<img src="https://static.igem.org/mediawiki/2017/c/c4/T--Munich--Results_20170803_Max_Gel.png"> | <img src="https://static.igem.org/mediawiki/2017/c/c4/T--Munich--Results_20170803_Max_Gel.png"> | ||
<ul> | <ul> | ||
| Line 3,025: | Line 3,025: | ||
<div class="date">Monday 7th</div> | <div class="date">Monday 7th</div> | ||
| − | <ul class="myEntry cas13a"> <!-- | + | <ul class="myEntry cas13a"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Protein-Purifcation of Lbu and Lsh</b> <!-- | + | <b>Protein-Purifcation of Lbu and Lsh</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols"> - </a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols"> - </a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max, Ludwig, Teeradon <!-- | + | Participants: Max, Ludwig, Teeradon <!-- --> |
</li> | </li> | ||
| Line 3,042: | Line 3,042: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Inoculation of main cultures of Rosetta p2CT-His-MBP-Lbu_C13a_WT and Lsh in LB(Carb+Cm) <!-- | + | Inoculation of main cultures of Rosetta p2CT-His-MBP-Lbu_C13a_WT and Lsh in LB(Carb+Cm) <!--All--> |
</li> | </li> | ||
| Line 3,074: | Line 3,074: | ||
</ul> | </ul> | ||
<div class="date">Tuesday 8th</div> | <div class="date">Tuesday 8th</div> | ||
| − | <ul class="myEntry other"> <!-- | + | <ul class="myEntry other"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>First test with 3D printed 96-well plate</b> <!-- | + | <b>First test with 3D printed 96-well plate</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">-</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">-</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 3,091: | Line 3,091: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Tested different concentrations of fluorescein in a 3D printed 96 well plate on paper. <!-- | + | Tested different concentrations of fluorescein in a 3D printed 96 well plate on paper. <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 3,103: | Line 3,103: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
No light bleeding was observed in the experiment | No light bleeding was observed in the experiment | ||
</li> | </li> | ||
| Line 3,136: | Line 3,136: | ||
<div class="date">Wednesday 9th</div> | <div class="date">Wednesday 9th</div> | ||
| − | <ul class="myEntry cas13a"> <!-- | + | <ul class="myEntry cas13a"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Expression and Harvesting of Cas13a variants</b> <!-- | + | <b>Expression and Harvesting of Cas13a variants</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Protein Purification</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Protein Purification</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max, Ludwig <!-- | + | Participants: Max, Ludwig <!-- --> |
</li> | </li> | ||
| Line 3,153: | Line 3,153: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Harvested Cas13a Lbu overexpression. <!-- | + | Harvested Cas13a Lbu overexpression. <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 3,198: | Line 3,198: | ||
<div class="date">Thursday 10th</div> | <div class="date">Thursday 10th</div> | ||
| − | <ul class="myEntry cas13a"> <!-- | + | <ul class="myEntry cas13a"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>His Purification of Cas13a Lbu and Harvesting of Cas13a Lsh</b> <!-- | + | <b>His Purification of Cas13a Lbu and Harvesting of Cas13a Lsh</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">His-Purification</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">His-Purification</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max, Ludwig, Milica, Sven, Benedikt <!-- | + | Participants: Max, Ludwig, Milica, Sven, Benedikt <!-- --> |
</li> | </li> | ||
| Line 3,294: | Line 3,294: | ||
</ul> | </ul> | ||
<div class="date">Friday 11th</div> | <div class="date">Friday 11th</div> | ||
| − | <ul class="myEntry cas13a"> <!-- | + | <ul class="myEntry cas13a"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Reverse HIS purification and SEC of Cas13a Lbu</b> <!-- | + | <b>Reverse HIS purification and SEC of Cas13a Lbu</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">His purification and SEC protein purification</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">His purification and SEC protein purification</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Ludwig, Benedikt, Max, Sven <!-- | + | Participants: Ludwig, Benedikt, Max, Sven <!-- --> |
</li> | </li> | ||
| Line 3,311: | Line 3,311: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Incubation of cleaved Lbu sample with Ni-NTA beads (ca. 7 mL) (1 h, 4 °C) <!-- | + | Incubation of cleaved Lbu sample with Ni-NTA beads (ca. 7 mL) (1 h, 4 °C) <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 3,330: | Line 3,330: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
See Gel image of 14.8. for results. | See Gel image of 14.8. for results. | ||
</li> | </li> | ||
| Line 3,397: | Line 3,397: | ||
</ul> | </ul> | ||
<div class="date">Monday 14th</div> | <div class="date">Monday 14th</div> | ||
| − | <ul class="myEntry cas13a"> <!-- | + | <ul class="myEntry cas13a"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>SDS PAGE of Lbu purifcations</b> <!-- | + | <b>SDS PAGE of Lbu purifcations</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">SDS-PAGE</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">SDS-PAGE</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Ludwig <!-- | + | Participants: Ludwig <!-- --> |
</li> | </li> | ||
| Line 3,414: | Line 3,414: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | SDS-PAGE of Lbu purification from last week <!-- | + | SDS-PAGE of Lbu purification from last week <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 3,478: | Line 3,478: | ||
</ul> | </ul> | ||
<div class="date">Wednesday 16th</div> | <div class="date">Wednesday 16th</div> | ||
| − | <ul class="myEntry cas13a"> <!-- | + | <ul class="myEntry cas13a"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Cas13a preparation</b> <!-- | + | <b>Cas13a preparation</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">-</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">-</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Benedikt, Ludwig <!-- | + | Participants: Benedikt, Ludwig <!-- --> |
</li> | </li> | ||
| Line 3,495: | Line 3,495: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Concentration measurment of Lbu, Lsh and Lwa with Bradford essay <!-- | + | Concentration measurment of Lbu, Lsh and Lwa with Bradford essay <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 3,507: | Line 3,507: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
Concentration of proteins measured(µg/ml): | Concentration of proteins measured(µg/ml): | ||
<li> | <li> | ||
| Line 3,560: | Line 3,560: | ||
</li> | </li> | ||
</ul> | </ul> | ||
| − | <ul class="myEntry cas13a"> <!-- | + | <ul class="myEntry cas13a"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Protein-Expression of Lbu and Lwa</b> <!-- | + | <b>Protein-Expression of Lbu and Lwa</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Protein-Expression</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Protein-Expression</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Benedikt, Ludwig, Milica <!-- | + | Participants: Benedikt, Ludwig, Milica <!-- --> |
</li> | </li> | ||
| Line 3,577: | Line 3,577: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Expression of Rosetta Cas13a Lbu (3L 2xYT Medium, 16°C ON) <!-- | + | Expression of Rosetta Cas13a Lbu (3L 2xYT Medium, 16°C ON) <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 3,586: | Line 3,586: | ||
</ul> | </ul> | ||
<div class="date">Friday 18th</div> | <div class="date">Friday 18th</div> | ||
| − | <ul class="myEntry cas13a"> <!-- | + | <ul class="myEntry cas13a"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Harvesting and Purification of Cas13a Lbu and Lsh</b> <!-- | + | <b>Harvesting and Purification of Cas13a Lbu and Lsh</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Protein Purification</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Protein Purification</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Benedikt, Milica <!-- | + | Participants: Benedikt, Milica <!-- --> |
</li> | </li> | ||
| Line 3,603: | Line 3,603: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Harvesting of both expressions <!-- | + | Harvesting of both expressions <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 3,615: | Line 3,615: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
</li> | </li> | ||
| Line 3,640: | Line 3,640: | ||
</ul> | </ul> | ||
<div class="date">Monday 21st</div> | <div class="date">Monday 21st</div> | ||
| − | <ul class="myEntry cas13a"> <!-- | + | <ul class="myEntry cas13a"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Continuation of Cas13a purification</b> <!-- | + | <b>Continuation of Cas13a purification</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">His-Purification</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">His-Purification</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Benedikt, Milica, Max <!-- | + | Participants: Benedikt, Milica, Max <!-- --> |
</li> | </li> | ||
| Line 3,657: | Line 3,657: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Continued His purification of Cas13a Lbu <!-- | + | Continued His purification of Cas13a Lbu <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 3,671: | Line 3,671: | ||
<ul> | <ul> | ||
<li>As visible on the gel the elutions 1 and 2 contained the highest amount of protein and were pooled for the TEV cleavage.</li> | <li>As visible on the gel the elutions 1 and 2 contained the highest amount of protein and were pooled for the TEV cleavage.</li> | ||
| − | </ul> <!-- | + | </ul> <!----> |
</li> | </li> | ||
| Line 3,756: | Line 3,756: | ||
</ul> | </ul> | ||
<div class="date">Tuesday 22nd</div> | <div class="date">Tuesday 22nd</div> | ||
| − | <ul class="myEntry cas13a"> <!-- | + | <ul class="myEntry cas13a"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Reverse His purification of Cas13a Lbu</b> <!-- | + | <b>Reverse His purification of Cas13a Lbu</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">His purification</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">His purification</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max, Benedikt, Milica <!-- | + | Participants: Max, Benedikt, Milica <!-- --> |
</li> | </li> | ||
| Line 3,773: | Line 3,773: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Incubation (shaker!!) of cleaved Lbu sample with Ni-NTA beads (ca. 7 mL) (1 h, ice) <!-- | + | Incubation (shaker!!) of cleaved Lbu sample with Ni-NTA beads (ca. 7 mL) (1 h, ice) <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 3,792: | Line 3,792: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
</li> | </li> | ||
| Line 3,880: | Line 3,880: | ||
</ul> | </ul> | ||
<div class="date">Wednesday 23rd</div> | <div class="date">Wednesday 23rd</div> | ||
| − | <ul class="myEntry cas13a"> <!-- | + | <ul class="myEntry cas13a"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Cas13a purification</b> <!-- | + | <b>Cas13a purification</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Size Exclusion Chromatography</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Size Exclusion Chromatography</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Benedikt, Milica <!-- | + | Participants: Benedikt, Milica <!-- --> |
</li> | </li> | ||
| Line 3,897: | Line 3,897: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Measuring concentration after Reverse His Lbu <!-- | + | Measuring concentration after Reverse His Lbu <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 3,909: | Line 3,909: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
See SDS PAGE of 24.8. to see the SEC result. | See SDS PAGE of 24.8. to see the SEC result. | ||
</li> | </li> | ||
| Line 3,976: | Line 3,976: | ||
</ul> | </ul> | ||
<div class="date">Thursday 24th</div> | <div class="date">Thursday 24th</div> | ||
| − | <ul class="myEntry cas13a"> <!-- | + | <ul class="myEntry cas13a"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>SDS-Gel of SEC-Run from 23.8.</b> <!-- | + | <b>SDS-Gel of SEC-Run from 23.8.</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">SDS-PAGE</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">SDS-PAGE</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Chris <!-- | + | Participants: Chris <!-- --> |
</li> | </li> | ||
| Line 3,993: | Line 3,993: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | 10% SDS PAGE of SEC Run samples <!-- | + | 10% SDS PAGE of SEC Run samples <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 4,003: | Line 4,003: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
<img src="https://static.igem.org/mediawiki/2017/3/31/T--Munich--Results_20170824_Max_Gel.png"> | <img src="https://static.igem.org/mediawiki/2017/3/31/T--Munich--Results_20170824_Max_Gel.png"> | ||
<ul><li>The samples with the highest protein concentration were pooled and used for further experiments.</li></ul> | <ul><li>The samples with the highest protein concentration were pooled and used for further experiments.</li></ul> | ||
| Line 4,255: | Line 4,255: | ||
<div class="date">Tuesday 29th</div> | <div class="date">Tuesday 29th</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">PCR with Q5 Polymerase, PCR Cleanup Kit, Restriction digest and Ligation</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">PCR with Q5 Polymerase, PCR Cleanup Kit, Restriction digest and Ligation</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 4,275: | Line 4,275: | ||
</li> | </li> | ||
<li> | <li> | ||
| − | PCR of SPlit Uni C und Split Uni N with Q5 Polymerase <!-- | + | PCR of SPlit Uni C und Split Uni N with Q5 Polymerase <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 4,460: | Line 4,460: | ||
</ul> | </ul> | ||
<div class="date">Thursday 31st</div> | <div class="date">Thursday 31st</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">E. comp Transformation</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">E. comp Transformation</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 4,477: | Line 4,477: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | E. comp transformation of E. coli Turbo with 1 µl of overnight ligation of 27.8. <!-- | + | E. comp transformation of E. coli Turbo with 1 µl of overnight ligation of 27.8. <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 4,663: | Line 4,663: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Three lysis methods were used: sonifcation, lysozyme digestion and heat lysis in SDS. | + | Three lysis methods were used: sonifcation, lysozyme digestion and heat lysis in SDS. All |
</li> | </li> | ||
<li> | <li> | ||
| Line 4,711: | Line 4,711: | ||
<div class="date">Monday 4th</div> | <div class="date">Monday 4th</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">E. comp Transformation</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">E. comp Transformation</a> <!---->(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 4,728: | Line 4,728: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Plates showed no colonies as there was no SOC step. <!-- | + | Plates showed no colonies as there was no SOC step. <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 4,815: | Line 4,815: | ||
</ul> | </ul> | ||
<div class="date">Tuesday 5th</div> | <div class="date">Tuesday 5th</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Agarose-Gel</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Agarose-Gel</a> <!---->(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 4,832: | Line 4,832: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Ran a small 1.75% TAE-Agarose Gel at 150 V for 50 minutes and post stained with Sybr Gold <!-- | + | Ran a small 1.75% TAE-Agarose Gel at 150 V for 50 minutes and post stained with Sybr Gold <!--All--> |
</li> | </li> | ||
</ul> | </ul> | ||
| Line 4,838: | Line 4,838: | ||
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Overnight Ligation of Intein-Extein-Construct</b> <!-- | + | <b>Overnight Ligation of Intein-Extein-Construct</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Ligation of DNA-Fragments</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Ligation of DNA-Fragments</a> <!---->(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 4,853: | Line 4,853: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Overnight ligation in ice bath with psb1C3 and psb4A5 as backbones and a 3:1 molar excess of insert <!-- | + | Overnight ligation in ice bath with psb1C3 and psb4A5 as backbones and a 3:1 molar excess of insert <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 4,863: | Line 4,863: | ||
</ul> | </ul> | ||
| − | <ul class="myEntry other"> <!-- | + | <ul class="myEntry other"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Preparation of chemical competent Turbo cells</b> <!-- | + | <b>Preparation of chemical competent Turbo cells</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Preparation of chemical competent cells</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Preparation of chemical competent cells</a> <!---->(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 4,880: | Line 4,880: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Plated one aliquot as negative controll <!-- | + | Plated one aliquot as negative controll <!--All--> |
</li> | </li> | ||
| Line 4,887: | Line 4,887: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | No colonies on plates, therefore usefull competent cells. <!-- | + | No colonies on plates, therefore usefull competent cells. <!----> |
</li> | </li> | ||
| Line 4,983: | Line 4,983: | ||
<div class="date">Wednesday 6th</div> | <div class="date">Wednesday 6th</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Chem and electro Transformation</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Chem and electro Transformation</a> <!---->(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 5,000: | Line 5,000: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Transformation into chem and e. competent E.coli Turbo and DH5 alpha cells. <!-- | + | Transformation into chem and e. competent E.coli Turbo and DH5 alpha cells. <!--All--> |
</li> | </li> | ||
| Line 5,007: | Line 5,007: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | No colonies in the evening, therefore the plates were left at room temperature overnight. <!-- | + | No colonies in the evening, therefore the plates were left at room temperature overnight. <!----> |
</li> | </li> | ||
| Line 5,036: | Line 5,036: | ||
</ul> | </ul> | ||
<div class="date">Thursday 7th</div> | <div class="date">Thursday 7th</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>RNA Intein-Extein-Readout</b> <!-- | + | <b>RNA Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Colony PCR</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Colony PCR</a> <!---->(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 5,053: | Line 5,053: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Picked 128 colonies from plates of 06.09.17 <!-- | + | Picked 128 colonies from plates of 06.09.17 <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 5,132: | Line 5,132: | ||
<div class="date">Monday 11th</div> | <div class="date">Monday 11th</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols"> - </a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols"> - </a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 5,149: | Line 5,149: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Precultures of 16 colonies from plates of 06.09.17. <!-- | + | Precultures of 16 colonies from plates of 06.09.17. <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 5,182: | Line 5,182: | ||
<div class="date">Tuesday 12th</div> | <div class="date">Tuesday 12th</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Mini-Prep with Qiagen Kit and Sequencing Sample preparation </a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Mini-Prep with Qiagen Kit and Sequencing Sample preparation </a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 5,199: | Line 5,199: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Mini Prep of precultures from 11.09.17 <!-- | + | Mini Prep of precultures from 11.09.17 <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 5,209: | Line 5,209: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | psB4A5_C-Terminal 1-7 and 3-4 were positive.<!-- | + | psB4A5_C-Terminal 1-7 and 3-4 were positive.<!----> |
</li> | </li> | ||
</ul> | </ul> | ||
<div class="date">Wednesday 13th</div> | <div class="date">Wednesday 13th</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Restriction digest and PCR protocol with Q5 polymerase</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Restriction digest and PCR protocol with Q5 polymerase</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 5,231: | Line 5,231: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Send Samples psB4A5_C-Terminal 1-7 and 3-4 for sequencing with primer VF <!-- | + | Send Samples psB4A5_C-Terminal 1-7 and 3-4 for sequencing with primer VF <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 5,243: | Line 5,243: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
PsB4A5_C terminal 1-7 was sequenced completly and used for further cloning steps. | PsB4A5_C terminal 1-7 was sequenced completly and used for further cloning steps. | ||
</li> | </li> | ||
| Line 5,305: | Line 5,305: | ||
</ul> | </ul> | ||
<div class="date">Thursday 14th</div> | <div class="date">Thursday 14th</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Restriction digest, Ligation of DNA fragments, Gibbson-Assembly and E and chem Trafo</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Restriction digest, Ligation of DNA fragments, Gibbson-Assembly and E and chem Trafo</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 5,322: | Line 5,322: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | New digestion of unamplified N-Terminal gBlock and psB1C3 with SpeI-HF and EcoRI-Hf <!-- | + | New digestion of unamplified N-Terminal gBlock and psB1C3 with SpeI-HF and EcoRI-Hf <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 5,341: | Line 5,341: | ||
</ul> | </ul> | ||
<div class="date">Friday 15th</div> | <div class="date">Friday 15th</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Colony PCR</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Colony PCR</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 5,358: | Line 5,358: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Picked 32 colonies from plates of 14.09.17 <!-- | + | Picked 32 colonies from plates of 14.09.17 <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 5,370: | Line 5,370: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
<img src="https://static.igem.org/mediawiki/2017/5/54/T--Munich--Results_20170915_Max_Gel.png"> | <img src="https://static.igem.org/mediawiki/2017/5/54/T--Munich--Results_20170915_Max_Gel.png"> | ||
<ul><li>No positive colononies in the Col PCR.</li></ul> | <ul><li>No positive colononies in the Col PCR.</li></ul> | ||
| Line 5,437: | Line 5,437: | ||
</ul> | </ul> | ||
<div class="date">Monday 18th</div> | <div class="date">Monday 18th</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Q5-PCR and Restriction digestion and Col-PCR and e and chem comp Transformation</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Q5-PCR and Restriction digestion and Col-PCR and e and chem comp Transformation</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 5,454: | Line 5,454: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | New PCR of backbone with psB1A3-fw and psB1A3-rev. Sample was subsequently DpnI digested and purified <!-- | + | New PCR of backbone with psB1A3-fw and psB1A3-rev. Sample was subsequently DpnI digested and purified <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 5,467: | Line 5,467: | ||
</ul> | </ul> | ||
<div class="date">Tuesday 19th</div> | <div class="date">Tuesday 19th</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout and Biobrick Recloning</b> <!-- | + | <b>Intein-Extein-Readout and Biobrick Recloning</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Gibbson-Assembly and Ligation</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Gibbson-Assembly and Ligation</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max, Rob <!-- | + | Participants: Max, Rob <!-- --> |
</li> | </li> | ||
| Line 5,484: | Line 5,484: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Gel of Col-PCR from 18.09.17 <!-- | + | Gel of Col-PCR from 18.09.17 <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 5,496: | Line 5,496: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
<img src="https://static.igem.org/mediawiki/2017/4/43/T--Munich--Results_20170919_Max_Gel.png"> | <img src="https://static.igem.org/mediawiki/2017/4/43/T--Munich--Results_20170919_Max_Gel.png"> | ||
<ul><li>Sample 1-3 and 2-1 had the correct length, and therefore precultures of these colonies were started overnight.</li></ul> | <ul><li>Sample 1-3 and 2-1 had the correct length, and therefore precultures of these colonies were started overnight.</li></ul> | ||
| Line 5,579: | Line 5,579: | ||
</ul> | </ul> | ||
<div class="date">Wednesday 20th</div> | <div class="date">Wednesday 20th</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Miniprep with Qiagen Kit</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Miniprep with Qiagen Kit</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 5,596: | Line 5,596: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Minipreped overnight cultures from 1-3 and 2-1. <!-- | + | Minipreped overnight cultures from 1-3 and 2-1. <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 5,606: | Line 5,606: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | N-Terminal sample 1-3 was positive. <!-- | + | N-Terminal sample 1-3 was positive. <!----> |
</li> | </li> | ||
</ul> | </ul> | ||
| − | <ul class="myEntry other"> <!-- | + | <ul class="myEntry other"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Biobrick Recloning</b> <!-- | + | <b>Biobrick Recloning</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Col PCR and preparation of e comp. cells</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Col PCR and preparation of e comp. cells</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 5,627: | Line 5,627: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Colony PCR of 36 colonies from each of the 3 constructs His-Sumo-Lwa (Lwa(l)), Lwa(lwa(k) and His-Sumo-Lbu each in psB1C3. <!-- | + | Colony PCR of 36 colonies from each of the 3 constructs His-Sumo-Lwa (Lwa(l)), Lwa(lwa(k) and His-Sumo-Lbu each in psB1C3. <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 5,667: | Line 5,667: | ||
</ul> | </ul> | ||
<div class="date">Thurdsay 21st</div> | <div class="date">Thurdsay 21st</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Restriction digest, Gibbson-Assembly and Transformation</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Restriction digest, Gibbson-Assembly and Transformation</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 5,684: | Line 5,684: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Digestion of 1 µg of each backbone (psb1c3_N-term_uni and psb4A5_C-Term_uni with BBS1-HF to open them for Gibbson-Assembly <!-- | + | Digestion of 1 µg of each backbone (psb1c3_N-term_uni and psb4A5_C-Term_uni with BBS1-HF to open them for Gibbson-Assembly <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 5,707: | Line 5,707: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
</li> | </li> | ||
</ul> | </ul> | ||
| − | <ul class="myEntry other"> <!-- | + | <ul class="myEntry other"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Biobrick Recloning</b> <!-- | + | <b>Biobrick Recloning</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Col-PCR and Sample Sequencing</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Col-PCR and Sample Sequencing</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 5,728: | Line 5,728: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Send samples for sequencing <!-- | + | Send samples for sequencing <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 5,740: | Line 5,740: | ||
<div class="date">Monday 25th</div> | <div class="date">Monday 25th</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Restriction digest, PCR Cleanup Kit, Gibbson Assembly and Transformation</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Restriction digest, PCR Cleanup Kit, Gibbson Assembly and Transformation</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 5,758: | Line 5,758: | ||
<li> | <li> | ||
Redigest of 1 µg of each backbone with BBS1 and dephoshorilation according to protocol | Redigest of 1 µg of each backbone with BBS1 and dephoshorilation according to protocol | ||
| − | <!-- | + | <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 5,781: | Line 5,781: | ||
| − | <ul class="myEntry other"> <!-- | + | <ul class="myEntry other"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Biobrick Recloning</b> <!-- | + | <b>Biobrick Recloning</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Total</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Total</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 5,798: | Line 5,798: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | COlPCR and Gel of samples from trafo of Friday 22.09.17 => 30 samples (all non green) psB1C3_LBU <!-- | + | COlPCR and Gel of samples from trafo of Friday 22.09.17 => 30 samples (all non green) psB1C3_LBU <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 5,808: | Line 5,808: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
All negative. | All negative. | ||
</li> | </li> | ||
</ul> | </ul> | ||
<div class="date">Tuesday 26th</div> | <div class="date">Tuesday 26th</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">ColPCR, Agarosegel</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">ColPCR, Agarosegel</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max, Milica <!-- | + | Participants: Max, Milica <!-- --> |
</li> | </li> | ||
| Line 5,830: | Line 5,830: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | ColPCR of 12 colonies from N-term plate from 25.9.17 <!-- | + | ColPCR of 12 colonies from N-term plate from 25.9.17 <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 5,839: | Line 5,839: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
Samples 1-8 and 2-1 of the N-Terminus looked good => preculture for sequencing. | Samples 1-8 and 2-1 of the N-Terminus looked good => preculture for sequencing. | ||
</li> | </li> | ||
| Line 5,845: | Line 5,845: | ||
| − | <ul class="myEntry other"> <!-- | + | <ul class="myEntry other"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Biobrick Recloning</b> <!-- | + | <b>Biobrick Recloning</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Total</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Total</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 5,862: | Line 5,862: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | COlPCR and Gel of samples from trafo of Friday 22.09.17 => 30 samples (all non green) psB1C3_Lwa_kurz <!-- | + | COlPCR and Gel of samples from trafo of Friday 22.09.17 => 30 samples (all non green) psB1C3_Lwa_kurz <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 5,871: | Line 5,871: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
All negative. | All negative. | ||
</li> | </li> | ||
</ul> | </ul> | ||
<div class="date">Wednesday 27th</div> | <div class="date">Wednesday 27th</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Minipreparation and Sample Sequencing</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Minipreparation and Sample Sequencing</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 5,893: | Line 5,893: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Miniprep of psB1C3_Split_N_term_bGal 1-8 and 2-1 <!-- | + | Miniprep of psB1C3_Split_N_term_bGal 1-8 and 2-1 <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 5,902: | Line 5,902: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
Both sequences positive. | Both sequences positive. | ||
</li> | </li> | ||
| Line 5,930: | Line 5,930: | ||
</ul> | </ul> | ||
<div class="date">Thursday 28th</div> | <div class="date">Thursday 28th</div> | ||
| − | <ul class="myEntry other"> <!-- | + | <ul class="myEntry other"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Biobrick Recloning</b> <!-- | + | <b>Biobrick Recloning</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">ColPCR, ON culture</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">ColPCR, ON culture</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 5,947: | Line 5,947: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | ColPCR from samples of 22.09.17 <!-- | + | ColPCR from samples of 22.09.17 <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 6,017: | Line 6,017: | ||
</ul> | </ul> | ||
<div class="date">Friday 29th</div> | <div class="date">Friday 29th</div> | ||
| − | <ul class="myEntry other"> <!-- | + | <ul class="myEntry other"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Biobrick Recloning</b> <!-- | + | <b>Biobrick Recloning</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Mini-Prepatation</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Mini-Prepatation</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 6,037: | Line 6,037: | ||
</li> | </li> | ||
<li> | <li> | ||
| − | Lwa short send for sequencing with VF and VR <!-- | + | Lwa short send for sequencing with VF and VR <!--All--> |
</li> | </li> | ||
| Line 6,044: | Line 6,044: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
Lwa long positive. | Lwa long positive. | ||
</li> | </li> | ||
| Line 6,224: | Line 6,224: | ||
<div class="date">Wednesday 4th</div> | <div class="date">Wednesday 4th</div> | ||
| − | <ul class="myEntry other"> <!-- | + | <ul class="myEntry other"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Biobrick Recloning</b> <!-- | + | <b>Biobrick Recloning</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">-</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">-</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 6,241: | Line 6,241: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Send Lwa short colony 2-3 for sequencing with pseqLwa primer <!-- | + | Send Lwa short colony 2-3 for sequencing with pseqLwa primer <!--All--> |
</li> | </li> | ||
| Line 6,248: | Line 6,248: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
Full read with no mutations. | Full read with no mutations. | ||
</li> | </li> | ||
</ul> | </ul> | ||
<div class="date">Thursday 5th</div> | <div class="date">Thursday 5th</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Transformation of e. comp cells</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Transformation of e. comp cells</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 6,270: | Line 6,270: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Repeated Gibbson-Assembly for Lbu with 4:1 ratio<!-- | + | Repeated Gibbson-Assembly for Lbu with 4:1 ratio<!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 6,279: | Line 6,279: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
Empty plates => need to repeat PCR. | Empty plates => need to repeat PCR. | ||
| Line 6,496: | Line 6,496: | ||
</ul> | </ul> | ||
<div class="date">Monday 9th</div> | <div class="date">Monday 9th</div> | ||
| − | <ul class="myEntry cas13a"> <!-- | + | <ul class="myEntry cas13a"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Lbu-Recloning with Gibbson-Assembly</b> <!-- | + | <b>Lbu-Recloning with Gibbson-Assembly</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">PCR,Gibbson and Trafo in chem and e comp</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">PCR,Gibbson and Trafo in chem and e comp</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 6,513: | Line 6,513: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | PCR for Gibbson with Biobrick fwd and rev primer. <!-- | + | PCR for Gibbson with Biobrick fwd and rev primer. <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 6,525: | Line 6,525: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
No colonies | No colonies | ||
</li> | </li> | ||
| Line 6,556: | Line 6,556: | ||
</ul> | </ul> | ||
<div class="date">Tuesday 10th</div> | <div class="date">Tuesday 10th</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">PCR and Gibbson Assembly</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">PCR and Gibbson Assembly</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 6,573: | Line 6,573: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | PCR of Lbu and psB1C3 as backbone <!-- | + | PCR of Lbu and psB1C3 as backbone <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 6,583: | Line 6,583: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
</li> | </li> | ||
| Line 6,620: | Line 6,620: | ||
<div class="date">Wednesday 11th</div> | <div class="date">Wednesday 11th</div> | ||
| − | <ul class="myEntry other"> <!-- | + | <ul class="myEntry other"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Biobrick Recloning </b> <!-- | + | <b>Biobrick Recloning </b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Transformation of chem comp cells</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Transformation of chem comp cells</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 6,637: | Line 6,637: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Transformation of Turbo cells with Gibbson of 10.10.<!-- | + | Transformation of Turbo cells with Gibbson of 10.10.<!--All--> |
</li> | </li> | ||
| Line 6,731: | Line 6,731: | ||
<div class="date">Thursday 12th</div> | <div class="date">Thursday 12th</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Transformation of e. comp cells</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Transformation of e. comp cells</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 6,748: | Line 6,748: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Transformation of BL21 with psB4A5 C-Term and psB1C3 N-Term <!-- | + | Transformation of BL21 with psB4A5 C-Term and psB1C3 N-Term <!--All--> |
</li> | </li> | ||
| Line 6,755: | Line 6,755: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
No colonies for psB4A5 C-Term. | No colonies for psB4A5 C-Term. | ||
</li> | </li> | ||
| Line 6,761: | Line 6,761: | ||
| − | <ul class="myEntry other"> <!-- | + | <ul class="myEntry other"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Biobrick Recloning</b> <!-- | + | <b>Biobrick Recloning</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Col-PCR for Biobrick Recloning</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Col-PCR for Biobrick Recloning</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 6,778: | Line 6,778: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Col PCR of 6x N-Term in psB1C3, 16x C-Term in psB1C3, 14x Lbu in psB1C3 <!-- | + | Col PCR of 6x N-Term in psB1C3, 16x C-Term in psB1C3, 14x Lbu in psB1C3 <!--All--> |
</li> | </li> | ||
</ul> | </ul> | ||
| Line 6,784: | Line 6,784: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
All negative. | All negative. | ||
</li> | </li> | ||
| Line 6,834: | Line 6,834: | ||
</ul> | </ul> | ||
<div class="date">Friday 13th</div> | <div class="date">Friday 13th</div> | ||
| − | <ul class="myEntry other"> <!-- | + | <ul class="myEntry other"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Biobrick Recloning</b> <!-- | + | <b>Biobrick Recloning</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">PCR with Q5-Polymerase</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">PCR with Q5-Polymerase</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 6,851: | Line 6,851: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Repeated PCR of N and C-term part for Recloning <!-- | + | Repeated PCR of N and C-term part for Recloning <!--All--> |
</li> | </li> | ||
</ul> | </ul> | ||
| Line 6,885: | Line 6,885: | ||
</ul> | </ul> | ||
<div class="date">Saturday 14th</div> | <div class="date">Saturday 14th</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Restriction digest Gibbson-Assembly and Transformation</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Restriction digest Gibbson-Assembly and Transformation</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 6,902: | Line 6,902: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | DpnI digest of PCR from 31.10.17 <!-- | + | DpnI digest of PCR from 31.10.17 <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 6,917: | Line 6,917: | ||
</ul> | </ul> | ||
<div class="date">Sunday 15th</div> | <div class="date">Sunday 15th</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Transformation of e. comp cells</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Transformation of e. comp cells</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 6,934: | Line 6,934: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Repeated Transformation of Bl21 star with psB4A5-Split-Intein-C-Term <!-- | + | Repeated Transformation of Bl21 star with psB4A5-Split-Intein-C-Term <!--All--> |
</li> | </li> | ||
</ul> | </ul> | ||
| Line 6,940: | Line 6,940: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
No colonies.=> Transformation in other expression strains | No colonies.=> Transformation in other expression strains | ||
</li> | </li> | ||
| Line 6,946: | Line 6,946: | ||
<div class="date">Monday 16th</div> | <div class="date">Monday 16th</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Transformation of e. comp cells</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Transformation of e. comp cells</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 6,963: | Line 6,963: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Transformed all available Bl21 strains with psB4A5-C-Term <!-- | + | Transformed all available Bl21 strains with psB4A5-C-Term <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 6,975: | Line 6,975: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
No colonies for any BL21 strain. The Turbo cells grew fine. => Expression seems to be toxic. | No colonies for any BL21 strain. The Turbo cells grew fine. => Expression seems to be toxic. | ||
</li> | </li> | ||
| Line 7,012: | Line 7,012: | ||
</ul> | </ul> | ||
<div class="date">Tuesday 17th</div> | <div class="date">Tuesday 17th</div> | ||
| − | <ul class="myEntry readout"> <!-- | + | <ul class="myEntry readout"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Intein-Extein-Readout</b> <!-- | + | <b>Intein-Extein-Readout</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">-</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">-</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 7,029: | Line 7,029: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Midipreparation of psB4A5-C-Term with Macherey-Nagel Xtra Midi <!-- | + | Midipreparation of psB4A5-C-Term with Macherey-Nagel Xtra Midi <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 7,041: | Line 7,041: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
In vitro expression was not done, due to lack of time. | In vitro expression was not done, due to lack of time. | ||
</li> | </li> | ||
</ul> | </ul> | ||
| − | <ul class="myEntry other"> <!-- | + | <ul class="myEntry other"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Biobrick Recloning</b> <!-- | + | <b>Biobrick Recloning</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Q5-PCR, Gibbson-Assembly and Transformation</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Q5-PCR, Gibbson-Assembly and Transformation</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 7,063: | Line 7,063: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | PCR of all GFP degradation tag library variants with Biobrick sequence binding primers, for transfer from psB1A3 to psB1C3. <!-- | + | PCR of all GFP degradation tag library variants with Biobrick sequence binding primers, for transfer from psB1A3 to psB1C3. <!--All--> |
</li> | </li> | ||
<li> | <li> | ||
| Line 7,073: | Line 7,073: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
</li> | </li> | ||
| Line 7,107: | Line 7,107: | ||
</ul> | </ul> | ||
<div class="date">Wednesday 18th</div> | <div class="date">Wednesday 18th</div> | ||
| − | <ul class="myEntry other"> <!-- | + | <ul class="myEntry other"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Biobrick Recloning</b> <!-- | + | <b>Biobrick Recloning</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Overnight culture</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Overnight culture</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 7,124: | Line 7,124: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Picked 2 green colonies from each plate for overnight culture <!-- | + | Picked 2 green colonies from each plate for overnight culture <!--All--> |
</li> | </li> | ||
| Line 7,160: | Line 7,160: | ||
</ul> | </ul> | ||
<div class="date">Thursday 19th</div> | <div class="date">Thursday 19th</div> | ||
| − | <ul class="myEntry other"> <!-- | + | <ul class="myEntry other"> <!--, existing categories are Cas13a,readout,other--> |
<li class="myTitel"> | <li class="myTitel"> | ||
| − | <b>Biobrick Recloning</b> <!-- | + | <b>Biobrick Recloning</b> <!----> |
</li> | </li> | ||
<li class="myProtocol"> | <li class="myProtocol"> | ||
| − | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Sequencing</a> <!-- | + | Protocol: <a href="https://2017.igem.org/Team:Munich/Protocols">Sequencing</a> <!--(NOT the href=..., the "Total RNA Purific...")--> |
</li> | </li> | ||
<li class="myParticipants"> | <li class="myParticipants"> | ||
| − | Participants: Max <!-- | + | Participants: Max <!-- --> |
</li> | </li> | ||
| Line 7,177: | Line 7,177: | ||
<ul> | <ul> | ||
<li> | <li> | ||
| − | Send samples from degradation tag library for sequencing <!-- | + | Send samples from degradation tag library for sequencing <!--All--> |
</li> | </li> | ||
</ul> | </ul> | ||
| Line 7,183: | Line 7,183: | ||
<li class="myResults"> | <li class="myResults"> | ||
<h4>Results</h4> | <h4>Results</h4> | ||
| − | <!-- | + | <!----> |
pdt2-E 2 was positive | pdt2-E 2 was positive | ||
</li> | </li> | ||
Revision as of 19:45, 31 October 2017
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