Difference between revisions of "Team:UMaryland/protocols"

 
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+
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margin:auto;
 
margin:auto;
 
font-family: "Palatino Linotype", "Book Antiqua", Palatino, serif;
 
font-family: "Palatino Linotype", "Book Antiqua", Palatino, serif;
 +
font-size:18px;
 
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#cover {
 
#cover {
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margin:auto;
 
margin:auto;
 
padding:0px;
 
padding:0px;
−
background-image: url("https://static.igem.org/mediawiki/2015/9/97/Resultsq.png");
+
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;
 +
z-index:3;
 
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}
−
 
  
 
#bar{
 
#bar{
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height:200px;
 
height:200px;
 
position:absolute;
 
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−
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+
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text-shadow: 4px 4px 4px white;
 
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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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<div id='bar'>
 
<div id='bar'>
 
<p style="font-size:64px"><b>Protocols</b>
 
<p style="font-size:64px"><b>Protocols</b>
 +
</div>
 
</div>
 
</div>
  
 +
 +
<div id='yolo'>
 +
<div id='sidemenu'>
 +
<ul>
 +
 +
<li><a href="#Miniprep" class="smoothScroll"><span>Miniprep</span></a></li>
 +
 +
<li><a href="#Ligation" class="smoothScroll"><span>Ligation</span></a></li>
 +
 +
<li><a href="#Transformation" class="smoothScroll"><span>Transformation</span></a></li>
 +
 +
<li><a href="#3Assembly" class="smoothScroll"><span>3A Assembly</span></a></li>
 +
 +
<li><a href="#REDigest" class="smoothScroll"><span>RE Digest</span></a></li>
 +
 +
<li><a href="#Gibsonassembly" class="smoothScroll"><span>Gibson Assembly</span></a></li>
 +
 +
<li><a href="#PCR" class="smoothScroll"><span>PCR</span></a></li>
 +
 +
<li><a href="#Gelextract" class="smoothScroll"><span>Gel Extract</span></a></li>
 +
<li><a href="#FPLC" class="smoothScroll"><span>FPLC</span></a></li>
 +
</ul>
 
</div>
 
</div>
  
−
<div id='layer2'>
+
 
 
<div id='contentbox'>
 
<div id='contentbox'>
 +
<a name="Miniprep">
 
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;">
 
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;">
 
<b>Miniprep</b></a>
 
<b>Miniprep</b></a>
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<div id='contentbox'>
 
<div id='contentbox'>
 +
<a name="Ligation">
 
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;">
 
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;">
 
<b>Ligation</b></a>
 
<b>Ligation</b></a>
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<div id='contentbox'>
 
<div id='contentbox'>
 +
<a name="Transformation">
 
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;">
 
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;">
 
<b>Transformation</b></a>
 
<b>Transformation</b></a>
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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>
−
   <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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</div>
 
</div>
 
<div id='contentbox'>
 
<div id='contentbox'>
 +
<a name="3Assembly">
 
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;">
 
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;">
 
<b>3A Assembly</b></a>
 
<b>3A Assembly</b></a>
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<div id='contentbox'>
 
<div id='contentbox'>
 +
<a name="REDigest">
 
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;">
 
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;">
 
<b>RE Digest</b></a>
 
<b>RE Digest</b></a>
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<div id='contentbox'>
 
<div id='contentbox'>
 +
<a name="Gibsonassembly">
 
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;">
 
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;">
 
<b>Gibson Assembly</b></a>
 
<b>Gibson Assembly</b></a>
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<div id='contentbox'>
 
<div id='contentbox'>
 +
<a name="PCR">
 
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;">
 
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;">
 
<b>PCR</b></a>
 
<b>PCR</b></a>
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</div>
 
</div>
 
<div id='contentbox'>
 
<div id='contentbox'>
 +
<a name="Gelextract">
 
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;">
 
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;">
 
<b>Gel Extract</b></a>
 
<b>Gel Extract</b></a>
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Procedure:
 
Procedure:
 
<ul class="a">
 
<ul class="a">
−
   <li>- cut out gel portions with scalpel</li>
+
   <li>- Cut out gel portions with scalpel</li>
−
   <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>
−
   <li>- discard flow through</li>
+
   <li>- Discard flow through</li>
−
   <li>- wash with .75 mL buffer PE in column, centrifuge 1 minute</li>
+
   <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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</div>
 
</div>
 
<div id='contentbox'>
 
<div id='contentbox'>
 +
<a name="FPLC">
 
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;">
 
<p style="font-size:32px;text-align:left;font-family:Verdana, Geneva, sans-serif;">
 
<b>Fast Protein Liquid Chromatography</b></a>
 
<b>Fast Protein Liquid Chromatography</b></a>
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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>
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   <li>- Wash off excess cobalt with syringe</li>
 
   <li>- Wash off excess cobalt with syringe</li>
 
   <li>- Connect affinity column to FPLC system</li>
 
   <li>- Connect affinity column to FPLC system</li>
−
   <li>- Inject supernatan</li>
+
   <li>- Inject supernatant</li>
 
   <li>- Wash FPLC column with equilibration column (X2) with syringe</li>
 
   <li>- Wash FPLC column with equilibration column (X2) with syringe</li>
 
</ul>
 
</ul>

Latest revision as of 02:32, 19 September 2015