Home Protocols High Efficiency Transformation Protocol for 96-tube format (C2987U)

High Efficiency Transformation Protocol for 96-tube format (C2987U)

  1. Remove the number of tubes you need from the 96-tube plate and thaw on ice for 4-5 minutes.
  2. Carefully remove the strip of caps from each tube and keep them for further use.
  3. Add 1-5 μl containing 1 pg-100 ng of plasmid DNA to each cell mixture. Cover the tubes with caps. Carefully flick the tube 4-5 times to mix cells and DNA. Do not vortex.
  4. Place the mixture on ice for 30 minutes. Do not mix. 
  5. Heat shock the cells at exactly 42°C for exactly 30 seconds in a water bath. Do not mix.
  6. Place on ice for 2 minutes. Do not mix.
  7. Pipette 500 µl of room temperature SOC into the mixture.
  8. Place at 37°C for 60 minutes. Shaking is not necessary.
  9. Warm selection plates to 37°C.
  10. Mix the cells thoroughly by pipetting, then perform several 10-fold serial dilutions in SOC or LB.
  11. Spread 50-100 μl of each dilution onto a selection plate and incubate overnight at 37°C.