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Team:UFAM Brazil/Protocol6
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Latest revision as of 15:23, 16 October 2014
Heat-Shock Transformation | ||
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Reagents: 1. 50ul of chemically competent cells 2. 22,5ul of MgCl2 3. 1ul plasmid Steps: 1. Allow chemically competent cell thawed for 1 hour on ice. 2. Add to the cells 22,5ul of MgCl2 and 1ul of plasmid, leaving on ice for 30 minutes. 3. Submit a heat-shock of 42 °C for 45 seconds. 4. Incubate the cells on ice for 10 minutes. 5. Add 500ul of LB medium and incubated in the shaker at 37 ° C for 50 to 60minutes. 6. Subculturing in LB agar with antibiotic selection. Do not forget the negative control! | ||
| Protocols | ||
