Dahlquist:Transformation Protocol for E. coli Chemically Competent Cells

Transformation Protocol for Competent E. coli DH1 Cells prepared by [[Media:DahlquistLab_CompetentCellPreparation.doc | 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.