Team:Macquarie Australia/WetLab/Protocols/Heat

Heat Shock

• Obtain competent cells from -80^oC.
• 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 42^oC water bath for 30 seconds, then back on ice for 2 min.
• Add 200µl of SOC media to each tube, and incubate in the 37^oC 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 37^oC incubator.