Dahlquist:Transformation Protocol for E. coli Chemically Competent Cells

From OpenWetWare

Jump to: navigation, search

Transformation Protocol for Competent E. coli DH1 Cells prepared by this method

  1. Thaw one tube of competent cells on ice.
  2. Add appropriate amount of DNA (or sterile water for no DNA control) to 200 μL of competent cells.
  3. Incubate the cells on ice for 40 min.
  4. Heat shock the cells at 42°C for 90 sec.
  5. Incubate the cells on ice for 5 min.
  6. Add 800 μL of LB to each tube using sterile technique. Incubate the tubes at 37°C for 30-60 min.
  7. Spin down your cells for 30 sec. at 12,000 rpm in the microfuge.
  8. Remove ~900 μL of supernatant. Resuspend the cells by vortexing and plate the suspension on LB plates containing 100 μg/mL ampicillin. Incubate at 37°C overnight.
Personal tools