Team:MIT mge
From 2010.igem.org
(Difference between revisions)
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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> |
Revision as of 17:48, 16 October 2010
Bacterial Construction Protocol
Bacterial Experimental Protocol
Phage western blot
Mammalian Protocol
Microfluidic stress
Mammalian Protocol
Microfluidics Protocol
Genetic Engineering Protocol
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
|