User:Isis Trenchard/Notebook/BioE44 Stuff/2010/03/19: Difference between revisions
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==Making | ==Making electrocompetent cells protocol== | ||
* Modified for general | * Modified for general electrocompetent cell prep (not specifically for recombineering): PMID: 19180090 | ||
#Pick an isolated colony from an LB plate and grow overnight in 3–5 ml of LB at 37C. | #Pick an isolated colony from an LB plate and grow overnight in 3–5 ml of LB at 37C. | ||
#Next morning, add 0.5 ml of the culture to 25 ml of LB in a 250-ml flask and grow at 37C to an OD600 of 0.50–0.60. Transfer the culture to a 50-ml Falcon tube and spin at 6,000g in prechilled rotor for 10 min at 4C. | #Next morning, add 0.5 ml of the culture to 25 ml of LB in a 250-ml flask and grow at 37C to an OD600 of 0.50–0.60. Transfer the culture to a 50-ml Falcon tube and spin at 6,000g in prechilled rotor for 10 min at 4C. |
Revision as of 09:54, 19 March 2010
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Making electrocompetent cells protocol
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