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Preparation of calcium competent Escherichia coli and heat ...

Vol. 1:22-25. Preparation of calcium competent Escherichia coli and heat-shock transformation Chang, Angela Y., Chau, Vivian WY., Landas, Julius A., Pang, Yvonne Department of Microbiology and Immunology, University of British Columbia calcium chloride heat-shock transformation is a powerful molecular biology technique used to introduce foreign DNA into a host cell. The concept of the technique is to render cells competent using CaCl2 to allow for introduction of plasmid. Plasmids usually contain the gene(s) of interest in addition to selection and/or antibiotic resistance markers. The Hanahan or calcium chloride method is used to generate chemically competent cells.

Buffer preparation (1-2hrs) C1. Subculturing overnight culture (3-4hrs) D3. Incubation (12-June 2017 Volume 1 Undergraduate Methods Paper 24 • Resuspend each pellet with 20mL ice-cold 0.1M CaCl 2, incubate on ice for 30 minutes • Centrifuge at 4°C at 4000rpm for 10 minutes ...

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