Difference between revisions of "Team:UMaryland/protocols"

 
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position:fixed;
 
position:fixed;
 
float:left;
 
float:left;
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width: 135px;
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width:200px;
 
top:200px;
 
top:200px;
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}
 
}
  
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   border-color: #ffffff;
 
   border-color: #ffffff;
 
}
 
}
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#layer1 {
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overflow:hidden;
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#yolo {
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width:100%;
 
width:100%;
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padding:20px;
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background-image: -webkit-linear-gradient(top, #FFFFFF 0%, #41CC6B 50%, #FFFFFF 100%);
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min-height:42px;
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top:-220px;
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}
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#layer2 {
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position:relative;
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background-image: -webkit-linear-gradient(top, #FFFFFF 0%, #46DBF2 50%, #FFFFFF 100%);
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min-height:42px;
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top:-220px;
 
top:-220px;
 
}
 
}
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#layer3 {
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#contentbox{
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width: 70%;
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left:240px;
 
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background-image: -webkit-linear-gradient(top, #FFFFFF 0%, #E6AC39 50%, #FFFFFF 100%);
 
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min-height:42px;
 
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}
 
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#layer4 {
 
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}
 
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width: 80%;
 
 
max-width: 1900px;
 
max-width: 1900px;
 
min-width: 1000px;
 
min-width: 1000px;
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margin:auto;
 
margin:auto;
 
font-family: "Palatino Linotype", "Book Antiqua", Palatino, serif;
 
font-family: "Palatino Linotype", "Book Antiqua", Palatino, serif;
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font-size:18px;
 
}
 
}
 
#cover {
 
#cover {
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margin:auto;
 
margin:auto;
 
padding:0px;
 
padding:0px;
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background-image: url("https://static.igem.org/mediawiki/2015/9/97/Resultsq.png");
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background-image: url("https://static.igem.org/mediawiki/2015/e/ed/Notebookumd.jpeg");
 
background-size: 100% ;
 
background-size: 100% ;
 
background-repeat: no-repeat;
 
background-repeat: no-repeat;
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font-size:xx-large;
 
font-size:xx-large;
 
top:-200px;
 
top:-200px;
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z-index:3;
 
}
 
}
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#bar{
 
#bar{
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height:200px;
 
height:200px;
 
position:absolute;
 
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text-shadow: 4px 4px 4px white;
 
text-shadow: 4px 4px 4px white;
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z-index:3;
 
}
 
}
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ul.a {list-style-type: circle;font-size:18px;color:black}
 
ul.a {list-style-type: circle;font-size:18px;color:black}
 
</style>
 
</style>
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<p style="font-size:64px"><b>Protocols</b>
 
<p style="font-size:64px"><b>Protocols</b>
 
</div>
 
</div>
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</div>
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<div id='yolo'>
 
<div id='sidemenu'>
 
<div id='sidemenu'>
 
<ul>
 
<ul>
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<li><a href="#Gelextract" class="smoothScroll"><span>Gel Extract</span></a></li>
 
<li><a href="#Gelextract" class="smoothScroll"><span>Gel Extract</span></a></li>
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<li><a href="#FPLC" class="smoothScroll"><span>GFPLC</span></a></li>
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<li><a href="#FPLC" class="smoothScroll"><span>FPLC</span></a></li>
 
</ul>
 
</ul>
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</div>
 
 
</div>
 
</div>
  
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   <li>- Heat shock at 42°C fro 30 seconds</li>
 
   <li>- Heat shock at 42°C fro 30 seconds</li>
 
   <li>- Add 1 mL SOC media to the cells</li>
 
   <li>- Add 1 mL SOC media to the cells</li>
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   <li>- Incubate for 60 minutes at 37°</li>
+
   <li>- Incubate for 60 minutes at 37°C</li>
 
   <li>- Plate 200 µL </li>
 
   <li>- Plate 200 µL </li>
 
   <li>- Incubate at 37°C</li>
 
   <li>- Incubate at 37°C</li>
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<ul class="a">
 
<ul class="a">
 
   <li>- Cut out gel portions with scalpel</li>
 
   <li>- Cut out gel portions with scalpel</li>
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   <li>- weigh gel slice in colorless tube. Add 3 volumes of QG buffer to 1 volume of gel</li>
+
   <li>- Weigh gel slice in colorless tube. Add 3 volumes of QG buffer to 1 volume of gel</li>
 
   <li>- Incubate at 50 for 10 mins until gels slice dissolves. Mix by vortexing tube every 2-3 mins during incubation to help the gel dissolve  
 
   <li>- Incubate at 50 for 10 mins until gels slice dissolves. Mix by vortexing tube every 2-3 mins during incubation to help the gel dissolve  
 
  </li>
 
  </li>
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   <li>- Place a Q1Aquick spin column in a provided 2ml collection tube</li>
 
   <li>- Place a Q1Aquick spin column in a provided 2ml collection tube</li>
 
   <li>- Apply sample to spin column, centrifuge for 1 min to bind dna</li>
 
   <li>- Apply sample to spin column, centrifuge for 1 min to bind dna</li>
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   <li>- discard flow through</li>
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   <li>- Discard flow through</li>
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   <li>- wash with .75 mL buffer PE in column, centrifuge 1 minute</li>
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   <li>- Wash with .75 mL buffer PE in column, centrifuge 1 minute</li>
 
   <li>- Place column in a clean 1.5 microcentrifuge tube</li>
 
   <li>- Place column in a clean 1.5 microcentrifuge tube</li>
 
   <li>- 40 microliters DDH20 , let stand 1 minute, centrifuge 1 min</li>
 
   <li>- 40 microliters DDH20 , let stand 1 minute, centrifuge 1 min</li>
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Procedure:
 
Procedure:
 
<ul class="a">
 
<ul class="a">
−
   <li>- centrifuge cell lysate (balance with equal mass of water) for 60 minutes at 25000 rpm</li>
+
   <li>- Centrifuge cell lysate (balance with equal mass of water) for 60 minutes at 25000 rpm</li>
 
   <li>- Wash FPLC column with equilibration column (X2) with  syringe</li>
 
   <li>- Wash FPLC column with equilibration column (X2) with  syringe</li>
 
   <li>- Wash out FLPC system with equilibration buffer</li>
 
   <li>- Wash out FLPC system with equilibration buffer</li>

Latest revision as of 02:32, 19 September 2015