Team:Macquarie Australia/heatshock

pbanner

Heat Shock Transformation




  1. Obtain competent cells from -80oC.
  2. Defrost gently on ice. 100µl is sufficient for 2 transformations.
  3. Add 1-10µl of plasmid DNA/ ligation mix to each tube. Incubate on ice for 30 min.
  4. Heat shock in 42oC water bath for 30 seconds, then back on ice for 2 min.
  5. Add 200µl of SOC media to each tube, and incubate in the 37oC shaker for 1h.
  6. For each tube of cells, spread 50µl onto one LB plate with appropriate antibiotic, and 100µl onto a second plate, using aseptic technique. Place your plate upside-down in the 37oC incubator.