Home Protocols High Efficiency Transformation Protocol for 384-well format (C2987R)

High Efficiency Transformation Protocol for 384-well format (C2987R)

  1. Chill a metal 384-well block on ice.
  2. Remove the plate from -80°C freezer, and place in chilled metal 384-well block for 2 minutes to thaw the competent cells.
  3. Carefully pierce holes through the foil seal.
  4. Add 1 μl containing 1 pg-100 ng of plasmid DNA to the cell mixture using a multichannel pipette. Carefully swirl the tips to mix cells and DNA.
  5. Seal the plate with plate cover.
  6. Incubate the plate in the chilled metal block for 20 minutes.
  7. Heat shock the cells at exactly 42°C for exactly 15 seconds by transferring the plate to a pre-warmed thermal block or water bath.
  8. Place in the chilled metal block for 2 minutes.
  9. Pipette 20 μl of room temperature Outgrowth Medium 1.5 into each well.
  10. Place at 37°C for 60 minutes. Shaking is not necessary.
  11. Warm selection plates to 37°C.
  12. Mix the cells thoroughly by pipetting, then perform several 10-fold serial dilutions in SOC or LB.
  13. Spread 50-100 μl of each dilution onto a selection plate and incubate overnight at 37°C.