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Team:Macquarie Australia/WetLab/Protocols/Heat
From 2014.igem.org
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| - | <h3>Heat Shock</h3> | + | <h3>Heat Shock</h3><br/> |
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<li>Obtain competent cells from -80<sup>o</sup>C.</li> | <li>Obtain competent cells from -80<sup>o</sup>C.</li> | ||
Revision as of 05:01, 16 October 2014
Heat Shock
- Obtain competent cells from -80oC.
- Defrost gently on ice. 100µl is sufficient for 2 transformations.
- Add 1-10µl of plasmid DNA/ ligation mix to each tube. Incubate on ice for 5 min.
- Put the tubes in the 42oC water bath for 30 seconds, then back on ice for 2 min.
- Add 200µl of SOC media to each tube, and incubate in the 37oC shaker for 10 min or up to 1h (10min for purified plasmid or 1h for ligation mix).
- For each tube of cells, spread 50µl onto one LB plate with appropriate antibiotic, and 500µl onto a second plate, using aseptic technique. Place your plate upside-down in the 37oC incubator.
