Shefigem2010: Transformation

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Transformation of Chemically Competent E.coli

1.) A 100ul aliquot of chemically competent cells was defrosted on ice for 10-15 min.

2.) The plasmid DNA was added to the cells, 1ul of ng quantities, and the mixture was left on ice for 15 min.

3.) The cells were heat-shocked at 42°C for 2 min and left on ice for 2 min before 1ml LB liquid media was added and the cells were allowed to recover at 37°C for 1 hour.

4.) Cells were pelleted via centrifugation at 13,000 rpm for 1 min and once resuspended in 100ul LB, plated out upon LB agar, containing the appropriate selective antibiotic.