Dahlquist:Transformation Protocol for E. coli Chemically Competent Cells: Difference between revisions
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(New page: Transformation Protocol for Competent E. coli DH1 Cells # Thaw one tube of competent cells on ice. # Add appropriate amount of DNA (or sterile water for no DNA control) to 200 μL of comp...) |
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Transformation Protocol for Competent E. coli DH1 Cells | Transformation Protocol for Competent E. coli DH1 Cells prepared by [[Media:DahlquistLab_CompetentCellPreparation.doc | this method]] | ||
# Thaw one tube of competent cells on ice. | # Thaw one tube of competent cells on ice. | ||
Latest revision as of 15:07, 6 August 2009
Transformation Protocol for Competent E. coli DH1 Cells prepared by this method
- Thaw one tube of competent cells on ice.
- Add appropriate amount of DNA (or sterile water for no DNA control) to 200 μL of competent cells.
- Incubate the cells on ice for 40 min.
- Heat shock the cells at 42°C for 90 sec.
- Incubate the cells on ice for 5 min.
- Add 800 μL of LB to each tube using sterile technique. Incubate the tubes at 37°C for 30-60 min.
- Spin down your cells for 30 sec. at 12,000 rpm in the microfuge.
- 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.
