Team:UIUC-Illinois/Notebook/Protocols

From 2012.igem.org

(Difference between revisions)
Line 10: Line 10:
</head>
</head>
-
<body>
+
<script src="http://ajax.googleapis.com/ajax/libs/jquery/1.7.2/jquery.min.js"></script>
-
<div id="protocol-container">
+
<script>
-
<div id="protocolselection">
+
-
<h1><center>Select Protocol</center></h1>
+
-
<ul class="protocolclass1">
+
$(document).ready(function(){
-
<li>
+
    $('a').click(function () {
-
<a href="https://2012.igem.org/Team:UIUC-Illinois/Notebook/Protocols/Bootcamp">Bootcamp Protocols</a>
+
        var divname= this.name;
-
</li>
+
        $("#"+divname)
 +
.fadeIn(300)
 +
.siblings()
 +
.hide(0);
 +
    });
 +
});
-
<li>
+
</script>
-
<a href="https://2012.igem.org/Team:UIUC-Illinois/Notebook/Protocols/Subculturing">Subculturing Plates</a>
+
</head>
-
</li>
+
<body>
-
<li>
+
    <div id="protocol-container">
-
<a href="https://2012.igem.org/Team:UIUC-Illinois/Notebook/Protocols/Electrocompetence">Making Electrocompetent E.Coli</a>
+
            <div id="protocolmenu">
-
</li>
+
                    <li><a name="prot1" >Bootcamp Protocols</a></li>
 +
                    <li><a name="prot2" >Digestions</a></li>
 +
                    <li><a name="prot3" >Gel Purification</a></li>
 +
                    <li><a name="prot4" >Inoculation</a></li>
 +
                    <li><a name="prot5" >Ligations</a></li>
 +
                    <li><a name="prot6" >Making Electrocompetent E.Coli</a></li>
 +
                    <li><a name="prot7" >Making Electrophoresis Gels</a></li>
 +
                    <li><a name="prot8" >Making TAE Buffers</a></li>
 +
                    <li><a name="prot9" >Miniprep</a></li>
 +
                    <li><a name="prot10" >PCR Protocols</a></li>
 +
                    <li><a name="prot11" >Storage of Cells</a></li>
 +
                    <li><a name="prot12" >Subculturing Plates</a></li>
 +
                    <li><a name="prot13" >Transformation of E.Coli</a></li>
 +
            </div>
-
<li>
+
<div id="protocoldescription">
-
<a href="https://2012.igem.org/Team:UIUC-Illinois/Notebook/Protocols/TAE">Making TAE Buffers</a>
+
-
</li>
+
-
<li>
+
<div id="prot1" style="display:none">
-
<a href="https://2012.igem.org/Team:UIUC-Illinois/Notebook/Protocols/GelElectrophoresis">Making Electrophoresis Gels</a>
+
Content1
-
</li>
+
-
 
+
-
<li>
+
-
<a href="https://2012.igem.org/Team:UIUC-Illinois/Notebook/Protocols/GelPurification">Gel Purification</a>
+
-
</li>
+
-
 
+
-
<li>
+
-
<a href="https://2012.igem.org/Team:UIUC-Illinois/Notebook/Protocols/Inoculation">Inoculation</a>
+
-
</li>
+
-
 
+
-
<li>
+
-
<a href="https://2012.igem.org/Team:UIUC-Illinois/Notebook/Protocols/Miniprep">Miniprep</a>
+
-
</li>
+
-
 
+
-
<li>
+
-
<a href="https://2012.igem.org/Team:UIUC-Illinois/Notebook/Protocols/PCR">PCR Protocols</a>
+
-
</li>
+
-
 
+
-
<li>
+
-
<a href="https://2012.igem.org/Team:UIUC-Illinois/Notebook/Protocols/Digestions">Digestions</a>
+
-
</li>
+
-
 
+
-
<li>
+
-
<a href="https://2012.igem.org/Team:UIUC-Illinois/Notebook/Protocols/Ligations">Ligations</a>
+
-
</li>
+
-
 
+
-
<li>
+
-
<a href="https://2012.igem.org/Team:UIUC-Illinois/Notebook/Protocols/Transformation">Transformation of E.Coli</a>
+
-
</li>
+
-
 
+
-
<li>
+
-
<a href="https://2012.igem.org/Team:UIUC-Illinois/Notebook/Protocols/Storage">Storage of Cells</a>
+
-
</li>
+
-
</ul>
+
</div>
</div>
-
 
+
<div id="prot2" style="display:none">
-
<div id="protocoloverview">
+
Content1
-
<center><h1>Bootcamp Protocols</h1></center>
+
</div>
-
 
+
<div id="prot3" style="display:none">
-
<h2>Making LB for plates</h2> <br/>
+
Content1
-
To make 1 Liter of LB: <br/><br/>
+
</div>
-
1. Always use a 10:5:5 ratio of Tryptone to Yeast Extract to NaCl. <br/>
+
<div id="prot4" style="display:none">
-
2. Add dry ingredients first. <br/>
+
Content1
-
3. Use a 2L flask. <br/> <br/>
+
</div>
-
4. Add the following <br/><br/>
+
<div id="prot5" style="display:none">
-
- 10 g Tryptone <br/>
+
Content1
-
- 5 g Yeast Extract <br/>
+
</div>
-
- 5 g NaCl <br/>
+
<div id="prot6" style="display:none">
-
- 1.5 g Agar (NOT agarose!)<br/><br/>
+
Content1
-
4. Then add 1 L of MilliQ water. <br/>
+
</div>
-
5. Autoclave by total volume.<br/>
+
<div id="prot7" style="display:none">
-
6. Pour 25 mL on each plate (just enough to cover the bottom). <br/><br/>
+
Content1
-
 
+
</div>
-
<h2>Making Liquid Media </h2> <br/>
+
<div id="prot8" style="display:none">
-
1. Always use a 5-2.5-2.5 ratio of Tryptone to Yeast Extract to NaCl <br/>
+
Content1
-
2. Add dry ingredients first, then add MilliQ water <br/>
+
</div>
-
3. No Agar! <br/>
+
<div id="prot9" style="display:none">
-
4. Autoclave by total volume <br/><br/>
+
Content1
-
 
+
</div>
-
<h2>Making Glycerol Stock </h2> <br/>
+
<div id="prot10" style="display:none">
-
To make 400 mL of 10% glycerol you will need:<br/><br/>
+
Content1
-
 
+
</div>
-
-40 mL glycerol<br/>
+
<div id="prot11" style="display:none">
-
-360 mL of MilliQ water <br/><br/>
+
Content1
-
 
+
</div>
-
To make 400 mL of 20% glycerol you will need:<br/><br/>
+
<div id="prot12" style="display:none">
-
 
+
Content1
-
-80 mL glycerol<br/>  
+
</div>
-
-320 mL of MilliQ water <br/><br/>
+
<div id="prot13" style="display:none">
-
 
+
Content1
-
1. Use flasks and bottles before graduated cylinders (they take forever to mix!)<br/>
+
-
2. Need a stir plate and a large stir bar <br/>
+
-
3. Stir until mixture is homogeneous <br/>
+
-
4. Don’t autoclave! <br/><br/>
+
-
 
+
-
<h2>Making ddH2O</h2> <br/>
+
-
- Use the small bottles. <br/>
+
-
- Autoclave water by total volume. <br/><br/>
+
-
 
+
-
<h2>Autoclaving </h2><br/>
+
-
- Robert is the source of official help on all things about the Autoclave. <br/><br/>
+
-
1. Use the autoclave tape. <br/>
+
-
2. Look at the Betastar for settings. <br/>
+
-
3. Go by materials and total volume. <br/>
+
-
4. Make sure the autoclave tape has changed colour by the end of the cycle. Check conditions and settings then repeat steps 1-4 if tape has not changed colour. <br/><br/>
+
-
 
+
-
<h2>Ligation </h2><br/>
+
-
We used the <a href="http://ginkgobioworks.com/support/BioBrick_Assembly_Manual.pdf">Ginkgo bioworks protocol</a> <br/><br/>
+
-
1. Control: <br/><br/>
+
-
- 2 uL of Circular PSB1C3 plasmid 13 uL ddH2O <br/>
+
-
- 2 uL T4 ligase buffer<br/>
+
-
- 1 uL T4 ligase <br/><br/>
+
-
2. P&C: <br/><br/>
+
-
- 2 uL of Circular PSB1C3 plasmid<br/>
+
-
- 2 uL of PUF, 11 uL ddH2O<br/>
+
-
- 2 uL T4 ligase buffer<br/>
+
-
- 1 uL T4 ligase<br/><br/>
+
-
3. L&P:<br/><br/>
+
-
- 2 uL of linear PSB1C3 plasmid<br/>
+
-
- 2 uL of PUF<br/>
+
-
- 11 uL ddH2O<br/>
+
-
- 2 uL T4 ligase buffer<br/>
+
-
- 1 uL T4 ligase<br/><br/>
+
-
 
+
-
 
+
</div>
</div>
</div>
</div>
-
 
-
</body>
 

Revision as of 22:04, 6 June 2012

Header

Protocols

  • Bootcamp Protocols
  • Digestions
  • Gel Purification
  • Inoculation
  • Ligations
  • Making Electrocompetent E.Coli
  • Making Electrophoresis Gels
  • Making TAE Buffers
  • Miniprep
  • PCR Protocols
  • Storage of Cells
  • Subculturing Plates
  • Transformation of E.Coli

    • Retrieved from "http://2012.igem.org/Team:UIUC-Illinois/Notebook/Protocols"