Team:Washington/HEAT SHOCK CHEM TRANS
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- Remove and thaw (on ice) chemically competent cells, either 1.5ml tube or PCR tube.
- 1.5mL tube aliquots of cells contain approx. 205uL cells
- Pcr tubes aliquots contain 50uls
- Add 1-5µl DNA to 40-50µl of competent cells.
- ncubate on ice for 5 minutes (for higher transformation efficiency, do 30 min)
- Heat shock cells for 45 seconds at 42°C
- Let cells stand on ice for 2 minutes (or longer)
- Add 200uL2-1mL of LB and shake for 30 min at 37°C (for higher transformation efficiency, do 1 hr). This is called recovery.
- Plate 200uL cells on appropriate antibiotic plate1 and grow O/N in 37°C incubator
Tips
- Preheat plates in 37C before for better results (labeling and cell absorption).
- If using 300ul pcr tubes, omit shaking, incubate 37C.
NOTE: Do not use XL10 Gold Cells with Chlor Plates