Team:MIT mge
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- | <a href="https://2010.igem.org/Team:MIT_bconst"> | + | <dl id="nav"> |
- | <a href="https://2010.igem.org/Team:MIT_bexp">Bacterial | + | <dt><b>Bacterial Protocol</b></dt> |
- | + | <dd> | |
- | <a href="https://2010.igem.org/Team:MIT_mmethods"> | + | <ul> |
- | + | <li><a href="https://2010.igem.org/Team:MIT_bconst">Biobrick Construction</a></li> | |
- | + | <li><a href="https://2010.igem.org/Team:MIT_bexp">Bacterial Experiments</a></li> | |
- | <a href="https://2010.igem.org/Team: | + | </ul> |
- | <a href="https://2010.igem.org/Team: | + | </dd> |
+ | <dt><b>Mammalian Protocol</b></dt> | ||
+ | |||
+ | <dd> | ||
+ | <ul> | ||
+ | <li><a href="https://2010.igem.org/Team:MIT_mmethods">Microfluidics</a></li> | ||
+ | <li><a href="https://2010.igem.org/Team:MIT_mge">Genetic Engineering</a></li> | ||
+ | <li><a href="https://2010.igem.org/Team:MIT_gateway">Gateway Cloning</a></li> | ||
+ | |||
+ | </ul> | ||
+ | </dd> | ||
+ | <dt><b>Phage Protocol</b></dt> | ||
+ | |||
+ | <dd> | ||
+ | <ul> | ||
+ | <li><a href="https://2010.igem.org/Team:MIT_phageprot">Basic Protocol</a></li> | ||
+ | |||
+ | </ul> | ||
+ | </dd> | ||
+ | |||
+ | </dl> | ||
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+ | <tr><td><div class="bodybaby">mammalian genetic engineering protocol</div></td> | ||
<tr><td><br>The Mammalian team created lines of cells using lentiviruses for transfection.<br><br> | <tr><td><br>The Mammalian team created lines of cells using lentiviruses for transfection.<br><br> | ||
<div class="outline"> | <div class="outline"> | ||
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<li>5.0g Hepes</li></ul></li> | <li>5.0g Hepes</li></ul></li> | ||
<li>Add all components into a beaker and add 400 mL of Nanopure/bidistilled water. Dissolve and adjust pH exactly to 7.06 (wait 20 min. to be sure that the pH won't change anymore).</li> | <li>Add all components into a beaker and add 400 mL of Nanopure/bidistilled water. Dissolve and adjust pH exactly to 7.06 (wait 20 min. to be sure that the pH won't change anymore).</li> | ||
- | + | <li>Filter through a 0.2 um filter, aliquot into 15 mL plastic tubes (12mL/tube). Can be kept at -20 for a few months.</li></ul><br> | |
+ | <b class="bolded" id="cacl">Calcium Chloride Solution (50mL)</b> | ||
+ | <ul id="procedure"> | ||
+ | <li>5M CaCl2 (18.4g CaCl2*2H2O in 50mL)</li> | ||
+ | <li>Keep at +4°C</li></ul><br> | ||
+ | <b class="bolded" id="freeze">Freezing medium</b> | ||
+ | <ul id="procedure"> | ||
+ | <li>12mL FCS</li> | ||
+ | <li>15mL DMEM (no antibiotics, directly as supplied)</li> | ||
+ | <li>3mL DMSO</li> | ||
+ | <li>TOTAL: 30mL</li></ul><br> | ||
+ | <b class="bolded" id="dmem">Supplemented DMEM</b> | ||
+ | <ul id="procedure"> | ||
+ | <li>450mL DMEM +Glucose +glutamine</li> | ||
+ | <li>50mL FBS</li> | ||
+ | <li>5mL Pen/Strep</li> | ||
+ | <li>0.5mL Fungin</li> | ||
+ | <li>TOTAL: 500mL</li></ul> | ||
</td> | </td> |
Latest revision as of 16:54, 27 October 2010
mammalian genetic engineering protocol |
The Mammalian team created lines of cells using lentiviruses for transfection. |
Calcium Phosphate Transfection Beforehand:
Protocol
Filter and Concentrate Virus Protocol
Materials HBSS Buffer (500 mL)
Calcium Chloride Solution (50mL)
Freezing medium
Supplemented DMEM
|