Difference between revisions of "Team:Munich/Testflow"

 
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href="https://2017.igem.org/Template:Munich/CSS?action=raw&ctype=text/css" />
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<link rel="stylesheet" type="text/css"
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<link rel="stylesheet" type="text/css"
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<head>
 
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+
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+
margin-bottom: 10px;
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+
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+
 
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+
 
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    background-image: url("wiki_image_menu_ihpgold_low.gif");
 
    display: none;
 
    position: absolute;
 
    top: 64;
 
    height: 32px;
 
    width: 137px;
 
    z-index: 1;
 
}
 
.dropdown:hover .dropdown-contentIHPgold {
 
    background-image: url("wiki_image_menu_ihpgold_high.gif");
 
    height: 32px;
 
    width: 137px;
 
    display: block;
 
}
 
.dropdown-contentIHPgold a {
 
    background-image: url("wiki_image_menu_ihpgold_low.gif");
 
    height: 32px;
 
    width: 137px;
 
    padding: 0px 0px;
 
    text-decoration: none;
 
    display: block;
 
}
 
.dropdown-contentIHPgold a:hover {
 
    background-image: url("wiki_image_menu_ihpgold_high.gif");
 
    height: 32px;
 
    width: 137px;
 
}
 
 
 
 
.dropdown-contentPublicEngagement {
 
    background-image: url("wiki_image_menu_publicengagement_low.gif");
 
    display: none;
 
    position: absolute;
 
    top: 96;
 
    height: 32px;
 
    width: 137px;
 
    z-index: 1;
 
}
 
.dropdown:hover .dropdown-contentPublicEngagement {
 
    background-image: url("wiki_image_menu_publicengagement_high.gif");
 
    height: 32px;
 
    width: 137px;
 
    display: block;
 
}
 
.dropdown-contentPublicEngagement a {
 
    background-image: url("wiki_image_menu_publicengagement_low.gif");
 
    height: 32px;
 
    width: 137px;
 
    padding: 0px 0px;
 
    text-decoration: none;
 
    display: block;
 
}
 
.dropdown-contentPublicEngagement a:hover {
 
    background-image: url("wiki_image_menu_publicengagement_high.gif");
 
    height: 32px;
 
    width: 137px;
 
}
 
 
 
 
.dropdown-contentCollaborations {
 
    background-image: url("wiki_image_menu_collaborations_low.gif");
 
    display: none;
 
    position: absolute;
 
    top: 128;
 
    height: 32px;
 
    width: 137px;
 
    z-index: 1;
 
}
 
.dropdown:hover .dropdown-contentCollaborations {
 
    background-image: url("wiki_image_menu_collaborations_high.gif");
 
    height: 32px;
 
    width: 137px;
 
    display: block;
 
}
 
.dropdown-contentCollaborations a {
 
    background-image: url("wiki_image_menu_collaborations_low.gif");
 
    height: 32px;
 
    width: 137px;
 
    padding: 0px 0px;
 
    text-decoration: none;
 
    display: block;
 
}
 
.dropdown-contentCollaborations a:hover {
 
    background-image: url("https://static.igem.org/mediawiki/2017/1/1c/T--Munich--wiki_image_menu_teammembers_high.gif");
 
    height: 32px;
 
    width: 137px;
 
}
 
 
 
 
.dropdown-contentTeamMembers {
 
    background-image: url("https://static.igem.org/mediawiki/2017/2/2b/T--Munich--wiki_image_menu_teammembers_low.gif");
 
    display: none;
 
    position: absolute;
 
    top: 32;
 
    height: 32px;
 
    width: 137px;
 
    z-index: 1;
 
}
 
.dropdown:hover .dropdown-contentTeamMembers {
 
    background-image: url("https://static.igem.org/mediawiki/2017/1/1c/T--Munich--wiki_image_menu_teammembers_high.gif");
 
    height: 32px;
 
    width: 137px;
 
    display: block;
 
}
 
.dropdown-contentTeamMembers a {
 
    background-image: url("wiki_image_menu_teammembers_low.gif");
 
    height: 32px;
 
    width: 137px;
 
    padding: 0px 0px;
 
    text-decoration: none;
 
    display: block;
 
}
 
.dropdown-contentTeamMembers a:hover {
 
    background-image: url("wiki_image_menu_teammembers_high.gif");
 
    height: 32px;
 
    width: 137px;
 
}
 
 
 
 
.dropdown-contentSupervisors {
 
    background-image: url("wiki_image_menu_supervisors_low.gif");
 
    display: none;
 
    position: absolute;
 
    top: 64;
 
    height: 32px;
 
    width: 137px;
 
    z-index: 1;
 
}
 
.dropdown:hover .dropdown-contentSupervisors {
 
    background-image: url("wiki_image_menu_supervisors_high.gif");
 
    height: 32px;
 
    width: 137px;
 
    display: block;
 
}
 
.dropdown-contentSupervisors a {
 
    background-image: url("wiki_image_menu_supervisors_low.gif");
 
    height: 32px;
 
    width: 137px;
 
    padding: 0px 0px;
 
    text-decoration: none;
 
    display: block;
 
}
 
.dropdown-contentSupervisors a:hover {
 
    background-image: url("wiki_image_menu_supervisors_high.gif");
 
    height: 32px;
 
    width: 137px;
 
}
 
 
 
 
.dropdown-contentPIs {
 
    background-image: url("wiki_image_menu_pis_low.gif");
 
    display: none;
 
    position: absolute;
 
    top: 96;
 
    height: 32px;
 
    width: 137px;
 
    z-index: 1;
 
}
 
.dropdown:hover .dropdown-contentPIs {
 
    background-image: url("wiki_image_menu_pis_high.gif");
 
    height: 32px;
 
    width: 137px;
 
    display: block;
 
}
 
.dropdown-contentPIs a {
 
    background-image: url("wiki_image_menu_pis_low.gif");
 
    height: 32px;
 
    width: 137px;
 
    padding: 0px 0px;
 
    text-decoration: none;
 
    display: block;
 
}
 
.dropdown-contentPIs a:hover {
 
    background-image: url("wiki_image_menu_pis_high.gif");
 
    height: 32px;
 
    width: 137px;
 
}
 
 
 
 
.dropdown-contentCollaboratingInstitutions {
 
    background-image: url("wiki_image_menu_collaboratinginstitutions_low.gif");
 
    display: none;
 
    position: absolute;
 
    top: 128;
 
    height: 32px;
 
    width: 137px;
 
    z-index: 1;
 
}
 
.dropdown:hover .dropdown-contentCollaboratingInstitutions {
 
    background-image: url("wiki_image_menu_collaboratinginstitutions_high.gif");
 
    height: 32px;
 
    width: 137px;
 
    display: block;
 
}
 
.dropdown-contentCollaboratingInstitutions a {
 
    background-image: url("wiki_image_menu_collaboratinginstitutions_low.gif");
 
    height: 32px;
 
    width: 137px;
 
    padding: 0px 0px;
 
    text-decoration: none;
 
    display: block;
 
}
 
.dropdown-contentCollaboratingInstitutions a:hover {
 
    background-image: url("wiki_image_menu_collaboratinginstitutions_high.gif");
 
    height: 32px;
 
    width: 137px;
 
}
 
 
 
 
.dropdown-contentSponsors {
 
    background-image: url("wiki_image_menu_sponsors_low.gif");
 
    display: none;
 
    position: absolute;
 
    top: 160;
 
    height: 32px;
 
    width: 137px;
 
    z-index: 1;
 
}
 
.dropdown:hover .dropdown-contentSponsors {
 
    background-image: url("wiki_image_menu_sponsors_high.gif");
 
    height: 32px;
 
    width: 137px;
 
    display: block;
 
}
 
.dropdown-contentSponsors a {
 
    background-image: url("wiki_image_menu_sponsors_low.gif");
 
    height: 32px;
 
    width: 137px;
 
    padding: 0px 0px;
 
    text-decoration: none;
 
    display: block;
 
}
 
.dropdown-contentSponsors a:hover {
 
    background-image: url("wiki_image_menu_sponsors_high.gif");
 
    height: 32px;
 
    width: 137px;
 
}
 
 
 
 
.dropdown-contentAttributions {
 
    background-image: url("wiki_image_menu_attributions_low.gif");
 
    display: none;
 
    position: absolute;
 
    top: 192;
 
    height: 32px;
 
    width: 137px;
 
    z-index: 1;
 
}
 
.dropdown:hover .dropdown-contentAttributions {
 
    background-image: url("wiki_image_menu_attributions_high.gif");
 
    height: 32px;
 
    width: 137px;
 
    display: block;
 
}
 
.dropdown-contentAttributions a {
 
    background-image: url("wiki_image_menu_attributions_low.gif");
 
    height: 32px;
 
    width: 137px;
 
    padding: 0px 0px;
 
    text-decoration: none;
 
    display: block;
 
}
 
.dropdown-contentAttributions a:hover {
 
    background-image: url("wiki_image_menu_attributions_high.gif");
 
    height: 32px;
 
    width: 137px;
 
}
 
 
#menu{
 
padding: 0 !important;
 
}
 
 
.mydescription{
 
height: 8em;
 
  overflow: hidden;
 
    cursor: pointer;
 
    cursor: hand; /*IE callst it differently. */
 
    position: relative;
 
}
 
 
.mydescription:hover{
 
animation: expandCard 0.5s;
 
animation-fill-mode: forwards;
 
}
 
 
#HQ_page .mydescription:hover p {
 
  background: initial;
 
  -webkit-background-clip: initial;
 
  -webkit-text-fill-color: initial;
 
}
 
 
.mydescription:hover .threeDots-container{
 
display: none;
 
}
 
 
@keyframes expandCard{
 
to{
 
height:15em;
 
}
 
}
 
 
.threeDots-container >img{
 
  height: auto;
 
  width: 50px;
 
  position: relative;
 
  left: -50%;
 
  top: -80%;
 
 
}
 
 
.threeDots-container{
 
  position: absolute;
 
  left: 50%;
 
  top: 80%;
 
 
 
} /*The container is there to make sure the threeDots picture is centered and on to bottom of the container (mydescription). No idea how or why this works. */
 
 
#HQ_page .mydescription >p{
 
  background: -webkit-linear-gradient(rgb(0,0,0),rgb(230,230,230) 50%, rgb(255,255,255));
 
  -webkit-background-clip: text;
 
  -webkit-text-fill-color: transparent;
 
  /*color: rgb(147,149,152);*/
 
  height: 15em;
 
  font-size: 1em;
 
}
 
 
.image > img{
 
  height: 200px !important;
 
}
 
 
.cards{
 
  align-items: flex-start;
 
}
 
 
 
</style>
 
</style>
</head><body>
+
</head></html>
 +
{{Munich/Menu}}
 +
<html>
 +
<body>
 
<table width=100% height=100% cellpadding=0 cellspacing=0 border=0>
 
<table width=100% height=100% cellpadding=0 cellspacing=0 border=0>
 
<!-- Bar -->
 
<tr><td id="menu" height="32" bgcolor=#51A7f9></td><td bgcolor=#51A7f9 height="32" width="960" align=center>
 
<div class="dropdown">
 
  <a href="index"><button class="dropbtnCascAid"><img src="overlay.png"></button></a>
 
  <div class="dropdown-content">
 
  </div>
 
</div><div class="dropdown">
 
  <a href="project" target=content><button class="dropbtnProject"><img src="overlay.png"></button></a>
 
  <div class="dropdown-contentDescription">
 
    <a href="description" target=content></a>
 
  </div>
 
  <div class="dropdown-contentDesign">
 
    <a href="design" target=content></a>
 
  </div>
 
  <div class="dropdown-contentProofofConcept">
 
    <a href="proofofconcept" target=content></a>
 
  </div>
 
  <div class="dropdown-contentDemonstration">
 
    <a href="demonstration" target=content></a>
 
  </div>
 
  <div class="dropdown-contentApplication">
 
    <a href="application" target=content></a>
 
  </div>
 
  <div class="dropdown-contentFinalResults">
 
    <a href="finalresults" target=content></a>
 
  </div>
 
  <div class="dropdown-contentEntrepreneurship">
 
    <a href="entrepreneurship" target=content></a>
 
  </div>
 
  <div class="dropdown-contentAchievements">
 
    <a href="achievements" target=content></a>
 
  </div>
 
</div><div class="dropdown" valign=center>
 
  <a href="wetlab" target=content><button class="dropbtnWetlab" valign=center><img src="overlay.png"></button></a>
 
  <div class="dropdown-contentProtocolsMethods">
 
    <a href="protocolsmethods" target=content></a>
 
  </div>
 
  <div class="dropdown-contentMaterials">
 
    <a href="materials" target=content></a>
 
  </div>
 
  <div class="dropdown-contentLabJournal">
 
    <a href="labjournal" target=content></a>
 
  </div>
 
  <div class="dropdown-contentSafety">
 
    <a href="safety" target=content></a>
 
  </div>
 
  <div class="dropdown-contentParts">
 
    <a href="parts" target=content></a>
 
  </div>
 
  <div class="dropdown-contentInterlab">
 
    <a href="interlab" target=content></a>
 
  </div>
 
</div><div class="dropdown">
 
  <a href="model" target=content><button class="dropbtnModel"><img src="overlay.png"></button></a>
 
  <div class="dropdown-content">
 
  </div>
 
</div><div class="dropdown">
 
  <a href="softhardware" target=content><button class="dropbtnSoftHardware"><img src="overlay.png"></button></a>
 
  <div class="dropdown-contentSoftware">
 
    <a href="software" target=content></a>
 
  </div>
 
  <div class="dropdown-contentHardware">
 
    <a href="hardware" target=content></a>
 
  </div>
 
</div><div class="dropdown">
 
  <a href="hp" target=content><button class="dropbtnHP"><img src="overlay.png"></button></a>
 
  <div class="dropdown-contentHPSilver">
 
    <a href="hpsilver" target=content></a>
 
  </div>
 
  <div class="dropdown-contentIHPgold">
 
    <a href="ihpgold" target=content></a>
 
  </div>
 
  <div class="dropdown-contentPublicEngagement">
 
    <a href="publicengagement" target=content></a>
 
  </div>
 
  <div class="dropdown-contentCollaborations">
 
    <a href="collaborations" target=content></a>
 
  </div>
 
</div><div class="dropdown">
 
  <a href="Team" target=content><button class="dropbtnTeam"><img src="overlay.png"></button></a>
 
  <div class="dropdown-contentTeamMembers">
 
    <a href="teammembers" target=content></a>
 
  </div>
 
  <div class="dropdown-contentSupervisors">
 
    <a href="supervisors" target=content></a>
 
  </div>
 
  <div class="dropdown-contentPIs">
 
    <a href="pis" target=content></a>
 
  </div>
 
  <div class="dropdown-contentCollaboratingInstitutions">
 
    <a href="collaboratinginstitutions" target=content></a>
 
  </div>
 
  <div class="dropdown-contentSponsors">
 
    <a href="sponsors" target=content></a>
 
  </div>
 
  <div class="dropdown-contentAttributions">
 
    <a href="attributions" target=content></a>
 
  </div>
 
</div>
 
</td><td height="32" bgcolor=#51A7f9></td></tr>
 
 
<!-- Content -->
 
<!-- Content -->
<tr><td width="100%" colspan=3>
+
<tr><td width="100%" colspan=4>
 
<table width=100% height=100% cellpadding=0 cellspacing=0 border=0>
 
<table width=100% height=100% cellpadding=0 cellspacing=0 border=0>
 
<tr>
 
<tr>
Line 1,152: Line 85:
 
<!-- Head End -->
 
<!-- Head End -->
 
<!-- Content Begin -->
 
<!-- Content Begin -->
<img width="960" src="https://static.igem.org/mediawiki/2017/a/ab/T--Munich--team-picture.png" style="margin-bottom: 15px">
+
<img id="TopPicture" width="960" src="https://static.igem.org/mediawiki/2017/0/04/T--Munich--FrontPagePictrues_FinalResults.jpg">
<table width="900" border=0 cellspacing=0 cellpadding=10>
+
<table width="960" border=0 cellspacing=0 cellpadding=10>
<tr><td colspan=3 align=left valign=center>
+
<tr>
<font size=7 color=#444444><b>Student</b></font>
+
<td width=160></td>
</td></tr>
+
<td width=160></td>
<tr><td colspan=3 align=center valign=center>
+
<td width=160></td>
<div class="ui cards centered stackable">
+
<td width=160></td>
      <div  class="ui card">
+
<td width=160></td>
          <div class="image">
+
<td width=160></td>
            <img src="https://static.igem.org/mediawiki/2017/c/cb/T--Munich--ProfilePicture_Pattrick.jpg">
+
</tr>
          </div>
+
<tr><td colspan=6 align=left valign=center>
        <div class="content">
+
<font size=7 color=#51a7f9><b style="color: #51a7f9">Final Results</b></font>
          <p class="header">Pattrick Wilke</p>
+
          <div class="meta">
+
            <span class="date">Masters Biochemistry (TUM)</span>
+
          </div>
+
          <div class="description" >
+
          <div class="mydescription" id="Pattrick" >
+
            <div class="threeDots-container">
+
              <img alt ="three dots" src="https://static1.squarespace.com/static/5654cf16e4b00b463710ff91/t/56568104e4b0509ba9df9296/1448509701389/?format=750w">
+
            </div>
+
            <p>
+
            He is currently in the Master in Biochemistry at TUM and is part of our Wet Lab Team. After spending too much time in the lab he has developed some side effects: he turned into a pipetting-robot and started to perceive reality in terms of incubation times. While he waits for the experiments to be done, he likes to play table football with Julian (you will know about him soon). Also, his laugh is very contagious.
+
            </p>
+
          </div>
+
          </div>
+
        </div>
+
      </div>
+
      <div class="ui card">
+
          <div class="image">
+
            <img src="https://static.igem.org/mediawiki/2017/e/e1/T--Munich--ProfilePicture_Chris.jpg">
+
          </div>
+
        <div class="content">
+
          <p class="header">Christoph Neumayer</p>
+
          <div class="meta">
+
            <span class="date">Bachelor Molecular Biotechnology (TUM)</span>
+
          </div>
+
          <div class="description" >
+
          <div class="mydescription" id="Christoph" >
+
            <div class="threeDots-container">
+
              <img alt ="three dots" src="https://static1.squarespace.com/static/5654cf16e4b00b463710ff91/t/56568104e4b0509ba9df9296/1448509701389/?format=750w">
+
            </div>
+
            <p>
+
            He is in his 4th semester in the Bachelor in Molecular Biotechnology at TUM. In his free time, he likes to make jokes about the other team members and he plays pool billiard on a professional level just to complete his geekiness. He spends most of his time at the lab and is one of our main guys in the Wet Lab Team. That explains why our food stocks are eaten mostly by him!
+
            </p>
+
          </div>
+
          </div>
+
        </div>
+
      </div>
+
      <div class="ui card">
+
          <div class="image">
+
            <img src="Profile_Pictures/Benedikt.jpg">
+
          </div>
+
        <div class="content">
+
          <p class="header">Benedikt Dürr</p>
+
          <div class="meta">
+
            <span class="date">Bachelor Biology (LMU)</span>
+
          </div>
+
          <div class="description" >
+
          <div class="mydescription" id="Benedikt" >
+
            <div class="threeDots-container">
+
              <img alt ="three dots" src="https://static1.squarespace.com/static/5654cf16e4b00b463710ff91/t/56568104e4b0509ba9df9296/1448509701389/?format=750w">
+
            </div>
+
            <p>
+
            Our youngest member and official paparazzi of Team Munich 2017. He is currently in the 4th semester of the Bachelor in Biology at LMU and begins to find the scientist in himself. However, this doesn’t hold him back from doing artsy stuff besides being in the lab. For that reason, he is in charge of the media and photography in our team. He designed not only our awesome project logo but also learned how to make videos just for the sake of the team!
+
            </p>
+
          </div>
+
          </div>
+
        </div>
+
      </div>
+
      <div class="ui card">
+
          <div class="image">
+
            <img src="https://static.igem.org/mediawiki/2017/d/d6/T--Munich--ProfilePicture_Sven.jpg">
+
          </div>
+
        <div class="content">
+
          <p class="header">Sven Klumpe</p>
+
          <div class="meta">
+
            <span class="date">Master Biochemistry (TUM)</span>
+
          </div>
+
          <div class="description" >
+
          <div class="mydescription" id="Sven" >
+
            <div class="threeDots-container">
+
              <img alt ="three dots" src="https://static1.squarespace.com/static/5654cf16e4b00b463710ff91/t/56568104e4b0509ba9df9296/1448509701389/?format=750w">
+
            </div>
+
            <p>
+
            At first, he joined the team to do simulation experiments but he ended up going to the lab almost every day. Now he leaves the lab very late and works in both the Wet Lab and Hardware Team. That´s what passion for science does to you! He is currently in the 3rd semester of the Master in Biochemistry at TUM. He is a big music fan and occasionally jams in a local bar. So, if you need a musical break from the lab work, you can ask this guy.
+
            </p>
+
          </div>
+
          </div>
+
        </div>
+
      </div>
+
      <div  class="ui card">
+
          <div class="image">
+
            <img src="https://static.igem.org/mediawiki/2017/a/ae/T--Munich--ProfilePicture_Erika.jpg">
+
          </div>
+
        <div class="content">
+
          <p class="header">Erika Chacin De Leonardis</p>
+
          <div class="meta">
+
            <span class="date">Master Biology (LMU)</span>
+
          </div>
+
          <div class="description" >
+
          <div class="mydescription" id="Erika" >
+
            <div class="threeDots-container">
+
              <img alt ="three dots" src="https://static1.squarespace.com/static/5654cf16e4b00b463710ff91/t/56568104e4b0509ba9df9296/1448509701389/?format=750w">
+
            </div>
+
            <p>
+
            She is from Venezuela and is currently in the 2nd semester of the Master in Biology at LMU. A few years ago, she tried to build an iGEM team and now she made it into one. She is very curious and always ready to learn new stuff. In our group, she is part of the Wet Lab Team. Her darkest secret is her love for metal music. Contrary to popular beliefs, coming from the Caribbean has not prepared her for European pre-summer temperatures, cue her ever present folding fan.
+
            </p>
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          </div>
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          </div>
+
        </div>
+
      </div>
+
      <div class="ui card">
+
          <div class="image">
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            <img src="Profile_Pictures/Luwdig.jpg">
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          </div>
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        <div class="content">
+
          <p class="header">Luwdig Bauer</p>
+
          <div class="meta">
+
            <span class="date">Master Biochemistry (TUM)</span>
+
          </div>
+
          <div class="description" >
+
          <div class="mydescription" id="Ludwig" >
+
            <div class="threeDots-container">
+
              <img alt ="three dots" src="https://static1.squarespace.com/static/5654cf16e4b00b463710ff91/t/56568104e4b0509ba9df9296/1448509701389/?format=750w">
+
            </div>
+
            <p>
+
            He is our Model Member of Team Munich 2017. He appears in most of our photos and we are beginning to believe that he just lives at the lab! He is in charge of the Wet Lab Team and always brings the good spirits to the rest of the team. He is currently in the 2nd semester of the Master in Biochemistry at TUM. Also, the last Linkin Park albums made him sad.
+
            </p>
+
          </div>
+
          </div>
+
        </div>
+
      </div>
+
      <div class="ui card">
+
          <div class="image">
+
            <img src="https://static.igem.org/mediawiki/2017/d/d4/T--Munich--ProfilePicture_Max.jpg">
+
          </div>
+
        <div class="content">
+
          <p class="header">Max Honeman</p>
+
          <div class="meta">
+
            <span class="date">Master Biochemistry (TUM)</span>
+
          </div>
+
          <div class="description" >
+
          <div class="mydescription" id="Max" >
+
            <div class="threeDots-container">
+
              <img alt ="three dots" src="https://static1.squarespace.com/static/5654cf16e4b00b463710ff91/t/56568104e4b0509ba9df9296/1448509701389/?format=750w">
+
            </div>
+
            <p>
+
            He is in the 4th semester of the Master in Biochemistry at TUM. He wanted to participate in iGEM this year so much that he delayed his Master Thesis. As a member of the Wet Lab Team, he helps with the organization of experiments and likes to play around with DNA and proteins. We need to warn you: he is our most sarcastic member and people tend to be confused and disoriented by his jokes, but we think he is nice anyways
+
            </p>
+
          </div>
+
          </div>
+
        </div>
+
      </div>
+
      <div class="ui card">
+
          <div class="image">
+
            <img src="https://static.igem.org/mediawiki/2017/6/61/T--Munich--ProfilePicture_Rob.jpg">
+
          </div>
+
        <div class="content">
+
          <p class="header">Rob Strasser</p>
+
          <div class="meta">
+
            <span class="date">Master Biology (LMU)</span>
+
          </div>
+
          <div class="description" >
+
          <div class="mydescription" id="Rob" >
+
            <div class="threeDots-container">
+
              <img alt ="three dots" src="https://static1.squarespace.com/static/5654cf16e4b00b463710ff91/t/56568104e4b0509ba9df9296/1448509701389/?format=750w">
+
            </div>
+
            <p>
+
            He is currently in the 2nd semester of the Master in Biology at LMU and is a big Immunology aficionado. He is just what you picture of a Biology student: carefree, always smiling, and shoeless to be one with nature! Within the team, he is part of the Paperstrip Team. Also, he has a thing for alpacas.
+
            </p>
+
          </div>
+
          </div>
+
        </div>
+
      </div>
+
      <div class="ui card">
+
          <div class="image">
+
            <img src="https://static.igem.org/mediawiki/2017/c/c7/T--Munich--Dali.jpg">
+
          </div>
+
        <div class="content">
+
          <p class="header">Dawafuti Sherpa</p>
+
          <div class="meta">
+
            <span class="date">Master Biology (LMU)</span>
+
          </div>
+
          <div class="description" >
+
          <div class="mydescription" id="Dali" >
+
            <div class="threeDots-container">
+
              <img alt ="three dots" src="https://static1.squarespace.com/static/5654cf16e4b00b463710ff91/t/56568104e4b0509ba9df9296/1448509701389/?format=750w">
+
            </div>
+
            <p>
+
            She is from Nepal and keeps the train moving in our team (actually she is our caretaker!) She takes matters in her own hands before anyone knows we needed to do something. She is in both the Human Practices and Wet Lab Teams. Currently she is in the Master in Biology at LMU. Also, in her free time she enjoys doing tai chi and sleeping.
+
            </p>
+
          </div>
+
          </div>
+
        </div>
+
      </div>
+
      <div class="ui card">
+
          <div class="image">
+
            <img src="https://static.igem.org/mediawiki/2017/f/f2/T--Munich--ProfilePicture_Milica.jpg">
+
          </div>
+
        <div class="content">
+
          <p class="header">Milica Zivanic</p>
+
          <div class="meta">
+
            <span class="date">Bachelor Molecular Biotechnology (TUM)</span>
+
          </div>
+
          <div class="description" >
+
          <div class="mydescription" id="Milica" >
+
            <div class="threeDots-container">
+
              <img alt ="three dots" src="https://static1.squarespace.com/static/5654cf16e4b00b463710ff91/t/56568104e4b0509ba9df9296/1448509701389/?format=750w">
+
            </div>
+
            <p>
+
            She is from Serbia and is currently in the 4th Semester of the Bachelor of Molecular Biotechnology at TUM. She is also in the Wiki Team and helps with the experiments in the Wet Lab. In her free time, she likes to teach kickboxing as a way to forget about bad experiments. Also, she is learning Spanish to have a life full of drama like in the telenovelas she watched as a kid.
+
            </p>
+
          </div>
+
          </div>
+
        </div>
+
      </div>
+
      <div class="ui card">
+
          <div class="image">
+
            <img src="https://static.igem.org/mediawiki/2017/3/3e/T--Munich--ProfilePicture_Jorge.jpg">
+
          </div>
+
        <div class="content">
+
          <p class="header">Jorge Andres Quintero Serna</p>
+
          <div class="meta">
+
            <span class="date">Bachelor Computer Science (LMU)</span>
+
          </div>
+
          <div class="description" >
+
          <div class="mydescription" id="Jorge" >
+
            <div class="threeDots-container">
+
              <img alt ="three dots" src="https://static1.squarespace.com/static/5654cf16e4b00b463710ff91/t/56568104e4b0509ba9df9296/1448509701389/?format=750w">
+
            </div>
+
            <p>
+
            He is from Colombia and previously completed a Bachelor in Biology at LMU, but since he likes to study so much, he decided to enroll in a Bachelor in Informatics after that! Currently, he is in charge of the Wiki development for our team. He is also an expert in The Beatles and you can challenge him on a trivia battle! (PD: he is a huge fan of DNA synthesis).
+
            </p>
+
          </div>
+
          </div>
+
        </div>
+
      </div>
+
      <div class="ui card">
+
          <div class="image">
+
            <img src="https://static.igem.org/mediawiki/2017/4/49/T--Munich--ProfilePicture_Katzi.jpg">
+
          </div>
+
        <div class="content">
+
          <p class="header">Florian Katzmeier</p>
+
          <div class="meta">
+
            <span class="date">Master Biophysics (TUM)</span>
+
          </div>
+
          <div class="description" >
+
          <div class="mydescription" id="Katzi" >
+
            <div class="threeDots-container">
+
              <img alt ="three dots" src="https://static1.squarespace.com/static/5654cf16e4b00b463710ff91/t/56568104e4b0509ba9df9296/1448509701389/?format=750w">
+
            </div>
+
            <p>
+
            He is the physicist guy of our team! He is currently in the 2nd semester of the Master in Biophysics at TUM. He really doesn´t enjoy Wet Lab activities and he is the happiest when he is around wires and circuit boards. So is not a surprise that he is part of the Hardware team. He is a big fan of precision, we know that from his almost obsessive behavior when using Inkscape.
+
            </p>
+
          </div>
+
          </div>
+
        </div>
+
      </div>
+
      <div class="ui card">
+
          <div class="image">
+
            <img src="Profile_Pictures/Julian.jpg">
+
          </div>
+
        <div class="content">
+
          <p class="header">Julian Reinhard</p>
+
          <div class="meta">
+
            <span class="date">Bachelor Bioinformatics (LMU/TUM)</span>
+
          </div>
+
          <div class="description" >
+
          <div class="mydescription" id="Julian" >
+
            <div class="threeDots-container">
+
              <img alt ="three dots" src="https://static1.squarespace.com/static/5654cf16e4b00b463710ff91/t/56568104e4b0509ba9df9296/1448509701389/?format=750w">
+
            </div>
+
            <p>
+
            He previously completed a Bachelor in Molecular Biotechnology at TUM and now he is in the 6th semester of the Bachelor in Bioinformatics, a partner program between LMU and TUM. In the lab, he is exploring different procedures for processing RNA and likes to play around with 3D printing. He likes to keep Patrick busy as his personal pipetting-robot while plotting to overthrow him as the team´s best table football player. By avoiding social media, he hopes his secret scheme will go unnoticed, but no more!
+
            </p>
+
          </div>
+
          </div>
+
        </div>
+
      </div>
+
      <div class="ui card">
+
          <div class="image">
+
            <img src="https://static.igem.org/mediawiki/2017/c/cf/T--Munich--ProfilePicture_Florian.jpg">
+
          </div>
+
        <div class="content">
+
          <p class="header">Florian Rothfischer</p>
+
          <div class="meta">
+
            <span class="date">Master Biochemistry (TUM)</span>
+
          </div>
+
          <div class="description" >
+
          <div class="mydescription" id="Florian" >
+
            <div class="threeDots-container">
+
              <img alt ="three dots" src="https://static1.squarespace.com/static/5654cf16e4b00b463710ff91/t/56568104e4b0509ba9df9296/1448509701389/?format=750w">
+
            </div>
+
            <p>
+
            He just finished his Bachelor in Biochemistry at TUM and decided to join iGEM before starting his Master degree next winter. He is part of the Wet Lab team and is working on different approaches for read-outs. During incubation times or meetings, he likes to cook pasta for the rest of the team. He finds lab work to be relaxing and the soundtrack for his experiments is performed by his favorite band: Florence + The Machine.
+
            </p>
+
          </div>
+
          </div>
+
        </div>
+
      </div>
+
      <div class="ui card">
+
          <div class="image">
+
            <img src="https://static.igem.org/mediawiki/2017/f/f1/T--Munich--ProfilePicture_Igor.jpg">
+
          </div>
+
        <div class="content">
+
          <p class="header">Igor Styazhkin</p>
+
          <div class="meta">
+
            <span class="date">Bachelor Biochemistry (TUM)</span>
+
          </div>
+
          <div class="description" >
+
          <div class="mydescription" id="Igor" >
+
            <div class="threeDots-container">
+
              <img alt ="three dots" src="https://static1.squarespace.com/static/5654cf16e4b00b463710ff91/t/56568104e4b0509ba9df9296/1448509701389/?format=750w">
+
            </div>
+
            <p>
+
            He is from Russia and is currently in the 4th semester of the Bachelor in Biochemistry at TUM, but he already has a Master in Telecommunications. In our group, he is part of the PR and Wiki Teams. Also, he is an expert in taking pictures and videos of us for our Instagram stories without anyone noticing (so probably he is a very good spy). Although he is tall and has a deep voice, don´t be afraid! He is a very nice guy.
+
            </p>
+
          </div>
+
          </div>
+
        </div>
+
      </div>
+
      <div class="ui card">
+
          <div class="image">
+
            <img src="https://static.igem.org/mediawiki/2017/5/5a/T--Munich--ProfilePicture_Matthias.jpg">
+
          </div>
+
        <div class="content">
+
          <p class="header">Matthias Lenz</p>
+
          <div class="meta">
+
            <span class="date">Bachelor Biology (TUM)</span>
+
          </div>
+
          <div class="description" >
+
          <div class="mydescription" id="Matthias" >
+
            <div class="threeDots-container">
+
              <img alt ="three dots" src="https://static1.squarespace.com/static/5654cf16e4b00b463710ff91/t/56568104e4b0509ba9df9296/1448509701389/?format=750w">
+
            </div>
+
            <p>
+
            The guy whose career in science was predetermined by his hair. He is in the 4th semester of the Bachelor of Biology at TUM. Matthias is in charge of getting sponsors and is also part of the Human Practices Team. He enjoys bouldering and has a 34 hours Spotify playlist of 60s rock, that he claims to consist only of his most favorite songs.
+
            </p>
+
          </div>
+
          </div>
+
        </div>
+
      </div>
+
      <div class="ui card">
+
          <div class="image">
+
            <img src="https://static.igem.org/mediawiki/2017/9/96/T--Munich--ProfilePicture_Teeradon.jpg">
+
          </div>
+
        <div class="content">
+
          <p class="header">Teeradon Phlairaharn</p>
+
          <div class="meta">
+
            <span class="date">Bachelor Molecular Biotechnology (TUM)</span>
+
          </div>
+
          <div class="description" >
+
          <div class="mydescription" id="Teeradon" >
+
            <div class="threeDots-container">
+
              <img alt ="three dots" src="https://static1.squarespace.com/static/5654cf16e4b00b463710ff91/t/56568104e4b0509ba9df9296/1448509701389/?format=750w">
+
            </div>
+
            <p>
+
            He comes from Thailand and is currently in the 2nd semester of the Bachelor in Molecular Biotechnology at TUM. He is part of the Human Practices team and is now working on ways to connect with people from different backgrounds to integrate our project within the community. He is always smiling and in his free time you can find him at ice cream shops.
+
            </p>
+
          </div>
+
          </div>
+
        </div>
+
      </div>
+
</div>
+
</td></tr>  
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<tr><td colspan=2 align=right valign=center height=10>
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<br><br><br><center><hr></center>
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</td></tr>
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</table>
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<!-- Content End -->
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<table width=960 height=35 cellpadding=0 cellspacing=0 border=0><tr><td align=right valign=center>
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<table width=300 height=35 cellpadding=0 cellspacing=0 border=0><tr>
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<td align=center valign=center>
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<a href="https://www.en.uni-muenchen.de/index.html" target="_blank3" >
+
<img src="https://static.igem.org/mediawiki/2017/8/83/T--Munich--wiki_image_coop_lmu_low.svg" border=0 height="20" onmouseover="this.src='https://static.igem.org/mediawiki/2017/f/fd/T--Munich--wiki_image_coop_lmu_high.svg'" onmouseout="this.src='https://static.igem.org/mediawiki/2017/8/83/T--Munich--wiki_image_coop_lmu_low.svg'">
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</a>
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</td>
 
</td>
 +
</tr>
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<tr>
 +
<td style="background-color: #51a7f9;" colspan = 6 align="left">
 +
<ul class="menuList" id="menu">
 +
  <li><a href="/Team:Munich/Cas13a">Cas13a</a></li>
 +
  <li><a href="/Team:Munich/Readouts">Readouts</a></li>
 +
  <li><a href="/Team:Munich/Targets">Targets</a></li>
 +
  <li><a href="/Team:Munich/Detection Chip">Detection Chip</a></li>
 +
  <li><a href="/Team:Munich/Amplification">Amplification</a></li>
 +
  <li><a href="/Team:Munich/Biobrick">Biobrick</a></li>
 +
</ul> 
  
<td align=center valign=center>
+
</td>
<a href="https://www.tum.de/" target="_blank4" >
+
</tr>
<img src="https://static.igem.org/mediawiki/2017/a/ae/T--Munich--wiki_image_coop_tum_low.svg" border=0 height="20" onmouseover="this.src='https://static.igem.org/mediawiki/2017/1/1e/T--Munich--wiki_image_coop_tum_high.svg'" onmouseout="this.src='https://static.igem.org/mediawiki/2017/a/ae/T--Munich--wiki_image_coop_tum_low.svg'">
+
</a>
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</td>
+
  
<td align=center valign=center>
 
<a href="http://www.e14.ph.tum.de/en/home/" target="_blank5" >
 
<img src="https://static.igem.org/mediawiki/2017/a/a3/T--Munich--wiki_image_coop_e14_low.svg" border=0 height="25" onmouseover="this.src='https://static.igem.org/mediawiki/2017/7/72/T--Munich--wiki_image_coop_e14_high.svg'" onmouseout="this.src='https://static.igem.org/mediawiki/2017/a/a3/T--Munich--wiki_image_coop_e14_low.svg'">
 
</a>
 
</td>
 
  
<td align=center valign=center>
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<tr><td colspan=6 align=center valign=center>
<a href="http://www.biochem.mpg.de/en" target="_blank5" >
+
<h1>Overview</h1>
<img src="https://static.igem.org/mediawiki/2017/d/da/T--Munich--wiki_image_coop_maxplanck_low.svg" border=0 height="30" onmouseover="this.src='https://static.igem.org/mediawiki/2017/d/df/T--Munich--wiki_image_coop_maxplanck_high.svg'" onmouseout="this.src='https://static.igem.org/mediawiki/2017/d/da/T--Munich--wiki_image_coop_maxplanck_low.svg'">
+
<p>
</a>
+
We demonstrated that each of the modules of our platform (extraction, amplification and detection of pathogenic RNA) is functional, although we did not yet fully integrate all the modules into a final product.
</td>
+
</p>
  
<td align=center valign=center>
+
<h3>What worked</h3>
<a href="https://www.helmholtz-muenchen.de/" target="_blank5" >
+
  <ul class="listResults">
<img src="https://static.igem.org/mediawiki/2017/0/05/T--Munich--wiki_image_coop_helmholtzzentrum_low.svg" border=0 height="20" onmouseover="this.src='https://static.igem.org/mediawiki/2017/6/6b/T--Munich--wiki_image_coop_helmholtzzentrum_high.svg'" onmouseout="this.src='https://static.igem.org/mediawiki/2017/0/05/T--Munich--wiki_image_coop_helmholtzzentrum_low.svg'">
+
          <li><a href="/Team:Munich/Cas13a">Demonstrated functionality of Lbu and Lwa Cas13a.</a></li>
</a>
+
          <li><a href="/Team:Munich/Cas13a">Modelled the detection limit of our circuit and confirmed it experimentally (~10 nM RNA).</a></li>
 +
          <li><a href="/Team:Munich/Cas13a">Detected pathogen RNA sequence from <i> in vitro </i> and <i> in vivo </i> sources.</a></li>
 +
          <li><a href="/Team:Munich/Targets">Differentiated viral sequences from bacterial sequences.</a></li>
 +
          <li><a href="/Team:Munich/Readouts">Used RNase Alert and Spinach aptamer read-out circuits.</a></li>
 +
          <li><a href="/Team:Munich/Readouts">Used gold nanoparticles to detect general RNase activity.</a></li>
 +
          <li><a href="/Team:Munich/Detection">Detected RNA in bulk, on paper, and from lyophilized Cas13a.</a></li>
 +
          <li><a href="/Team:Munich/Detection">Constructed a functional fluorescence detector with high sensitivity and low production cost.</a></li>
 +
          <li><a href="/Team:Munich/Cas13a">Detected RNA in bulk, on paper, and from lyophilized Cas13a.</a></li>
 +
          <li><a href="/Team:Munich/Amplification">Amplified target with RPA and transcription on paper.</a></li>
 +
          <li><a href="/Team:Munich/Parts">Improved the biobrick BBa_K1319008 by adding a 6x His-tag and provided Cas13a Lwa as three different composite biobricks.</a></li>
 +
          <li><a href="/Team:Munich/Part_Collection">Characterized the GFP degradation tags and sent them as a part collection.</a></li>
 +
  </ul>
 +
<h3>What presented issues</h3>
 +
  <ul class="listResults">
 +
          <li><a href="/Team:Munich/Cas13a">Optimizing the purification protocol for Cas13a.</a></li>
 +
          <li><a href="/Team:Munich/Cas13a">Demonstrating functionality of Lsh Cas13a.</a></li>
 +
          <li><a href="/Team:Munich/Cas13a">Ruling out RNase contamination from heat-lysed in vivo samples.</a></li>
 +
          <li><a href="/Team:Munich/Targets">Detecting Q5 beta RNA.</a></li>
 +
          <li><a href="/Team:Munich/Targets">Reducing cross-talk between <i> E.coli </i> crRNA and <i> B.subtilitis </i> target RNA.</a></li>
 +
          <li><a href="/Team:Munich/Readouts">Developing colorimetric read-outs.</a></li>
 +
          <li><a href="/Team:Munich/Detection">Optimizing the lyophilization and stability of Cas13a.</a></li>
 +
          <li><a href="/Team:Munich/Amplification">Performing RPA and transcription on chip.</a></li>
 +
  </ul>
 
</td>
 
</td>
 +
</tr>
  
<td align=center valign=center>
+
<tr><td align=center valign=center colspan=3>
<a href="https://igem.org/Main_Page" target="_blank6" >
+
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+
<h1>Discussion</h1>
<img src="https://static.igem.org/mediawiki/2017/8/8b/T--Munich--wiki_image_sm_twitter_low.svg" border=0 height="25" onmouseover="this.src='https://static.igem.org/mediawiki/2017/c/cc/T--Munich--wiki_image_sm_twitter_high.svg'" onmouseout="this.src='https://static.igem.org/mediawiki/2017/8/8b/T--Munich--wiki_image_sm_twitter_low.svg'">
+
<p> 
</a>
+
Our project CascAID is a universal solution for low cost, point of care diagnostics of infectious diseases. Currently, the available diagnostic tools are based on PCR, antibodies or microbiological methods which all need trained personal and lab equipments. Therefore, these methods are cost and time consuming. This gives rise to the need of developing effective, affordable and portable devices.</p><p>
 +
In our project, we first successfully replicated the Cas13a-based detection of RNA pathogens that was demonstrated by Gootenberg et al. Although this result is not novel, we thoroughly characterized the target detection limit for different bacterial and viral targets, from <i> in vitro </i> and <i> in vivo </i> sources, and proved the possibility to discriminate between viruses and bacteria with high specificity. We laid the groundwork for colorimetric read-outs that will add another layer of amplification in our cascade detection (gold nanoparticles, intein-extein and ssDNA amplification). Those readouts should allow for a practical readability of the diagnosis by the user without the need of digital analysis. Additionally, their amplification scheme should also lower the detection limit of the Cas13a without the need for pre-amplification of the target.  </p><p>
 +
However, we worked in parallel on a scheme for amplifying the target using RPA and transcription. Although the reaction worked on paper, it did not work on chip due to the toxicity of the PDMS to the reaction.  
 +
We built a fluorescence detector with high sensitivity to cost ratio, and used it successfully to detect Cas13a activity. However, the product itself needs to be redesigned for market distribution: in general, a fluorescence detector is not necessarily user-friendly, the extraction of the RNA on chip needs to be optimized, and the costs of the whole product must be lowered. </p><p>
 +
Nevertheless, we are glad to have created a functional platform that allows the detection of nanomolar concentrations of pathogens within 30 minutes. With our modular approach, we have shown at least proof-of-concept results for each part, and are confident that no fundamental gap prevents our platform from being usable, only optimization.  
 +
</p>
 
</td>
 
</td>
 +
</tr>
  
<td align=center valign=center>
+
<tr><td colspan=6 align=center valign=center>
<a href="mailto:igem.munich@gmail.com">
+
<h1>Outlook</h1>
<img src="https://static.igem.org/mediawiki/2017/7/79/T--Munich--wiki_image_sm_mail_low.svg" border=0 height="25" onmouseover="this.src='https://static.igem.org/mediawiki/2017/9/93/T--Munich--wiki_image_sm_mail_high.svg'" onmouseout="this.src='https://static.igem.org/mediawiki/2017/7/79/T--Munich--wiki_image_sm_mail_low.svg'">
+
<p> 
</a>
+
We still have some project sections that we need to improve in the future. We have therefore listed the following points that need to be optimized below.
 +
</p>
 +
  <ul class="listResults">
 +
          <li><i> In vivo </i> heat lysis: During our experiments, we realized that the RNA extraction of <i> E. coli </i> using heat lysis is not always optimal for our experimental setup due to the fact that we have RNase contamination in the extracted RNA samples. Although our Cas13a cleavage assays are performed in presence RNase Inhibitor to suppress the activity of the RNases that could be present, we saw that the heat lysed samples show relatively higher fluorescence activity in comparison to the phenol chloroform extracted samples</li>
 +
          <li>RNA extraction and amplification: The RNA extraction from the <i> Bacillus subtilis </i> was particularly difficult in our case since <i> B. subtilis </i> is a gram positive, spore forming bacteria. Also the amount and the quality of the RNA extracted from the <i> B. subtilis </i> and <i> E.coli </i> cultures were sufficiently good. We therefore should find methods to improve either the RNA extraction protocol or use a better amplification steps after the extraction.</li>
 +
          <li>Cost of the chip: Now, the cost of our chip is less than 15 dollars per chip. We could still try to minimize the costs by reducing the chip size and making it fully recyclable. However, at industrial level one could potentially reduce the cost of the chip.</li>
 +
          <li>Lyophilization of Cas13a: We also figured out that the lyophilization protocol of the Cas13a has to be improved in order to make our paper chip portable and sustainable. We also tried drying the Cas13a with the tardigrade intrinsically disordered proteins (TDPs) from team Delft but still it wasn’t that effective as expected. Therefore, we have to integrate some better methods to lyophilize the Cas13a without losing its activity.</li>
 +
          <li>Readouts with color and amplification: The colorimetric readout is also something we need to work on and improve since we only managed to partially succeed with the colorimetric assays. We however think that it is possible to realize this using more elegant ways of RNA detection and this is something we could try in future.</li>
 +
          <li>Integration of all the modules of the platform: Although all our modules parts are functional, we were only able to integrate them partially. So, with more time, we believe that we can have a fully functional and integrated module system.  
 +
</li>
 +
  </ul>
 
</td>
 
</td>
 +
</tr>
  
<td align=center valign=center>
 
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<td align=center valign=center>
+
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Latest revision as of 19:24, 31 October 2017


Final Results

Overview

We demonstrated that each of the modules of our platform (extraction, amplification and detection of pathogenic RNA) is functional, although we did not yet fully integrate all the modules into a final product.

What worked

What presented issues

Discussion

Our project CascAID is a universal solution for low cost, point of care diagnostics of infectious diseases. Currently, the available diagnostic tools are based on PCR, antibodies or microbiological methods which all need trained personal and lab equipments. Therefore, these methods are cost and time consuming. This gives rise to the need of developing effective, affordable and portable devices.

In our project, we first successfully replicated the Cas13a-based detection of RNA pathogens that was demonstrated by Gootenberg et al. Although this result is not novel, we thoroughly characterized the target detection limit for different bacterial and viral targets, from in vitro and in vivo sources, and proved the possibility to discriminate between viruses and bacteria with high specificity. We laid the groundwork for colorimetric read-outs that will add another layer of amplification in our cascade detection (gold nanoparticles, intein-extein and ssDNA amplification). Those readouts should allow for a practical readability of the diagnosis by the user without the need of digital analysis. Additionally, their amplification scheme should also lower the detection limit of the Cas13a without the need for pre-amplification of the target.

However, we worked in parallel on a scheme for amplifying the target using RPA and transcription. Although the reaction worked on paper, it did not work on chip due to the toxicity of the PDMS to the reaction. We built a fluorescence detector with high sensitivity to cost ratio, and used it successfully to detect Cas13a activity. However, the product itself needs to be redesigned for market distribution: in general, a fluorescence detector is not necessarily user-friendly, the extraction of the RNA on chip needs to be optimized, and the costs of the whole product must be lowered.

Nevertheless, we are glad to have created a functional platform that allows the detection of nanomolar concentrations of pathogens within 30 minutes. With our modular approach, we have shown at least proof-of-concept results for each part, and are confident that no fundamental gap prevents our platform from being usable, only optimization.

Outlook

We still have some project sections that we need to improve in the future. We have therefore listed the following points that need to be optimized below.

  • In vivo heat lysis: During our experiments, we realized that the RNA extraction of E. coli using heat lysis is not always optimal for our experimental setup due to the fact that we have RNase contamination in the extracted RNA samples. Although our Cas13a cleavage assays are performed in presence RNase Inhibitor to suppress the activity of the RNases that could be present, we saw that the heat lysed samples show relatively higher fluorescence activity in comparison to the phenol chloroform extracted samples
  • RNA extraction and amplification: The RNA extraction from the Bacillus subtilis was particularly difficult in our case since B. subtilis is a gram positive, spore forming bacteria. Also the amount and the quality of the RNA extracted from the B. subtilis and E.coli cultures were sufficiently good. We therefore should find methods to improve either the RNA extraction protocol or use a better amplification steps after the extraction.
  • Cost of the chip: Now, the cost of our chip is less than 15 dollars per chip. We could still try to minimize the costs by reducing the chip size and making it fully recyclable. However, at industrial level one could potentially reduce the cost of the chip.
  • Lyophilization of Cas13a: We also figured out that the lyophilization protocol of the Cas13a has to be improved in order to make our paper chip portable and sustainable. We also tried drying the Cas13a with the tardigrade intrinsically disordered proteins (TDPs) from team Delft but still it wasn’t that effective as expected. Therefore, we have to integrate some better methods to lyophilize the Cas13a without losing its activity.
  • Readouts with color and amplification: The colorimetric readout is also something we need to work on and improve since we only managed to partially succeed with the colorimetric assays. We however think that it is possible to realize this using more elegant ways of RNA detection and this is something we could try in future.
  • Integration of all the modules of the platform: Although all our modules parts are functional, we were only able to integrate them partially. So, with more time, we believe that we can have a fully functional and integrated module system.