IGEM:Peking/2007/Count:Transformation: Difference between revisions
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(New page: #Hold competent cells (from -80℃ refrigerator) on ice. #Gently mix ligation product (10-15 μL) with cells, then bath in ice for 30 min. #(Precisely!) Bath in 42℃ water for 90 sec. #Ho...) |
Qu Mingzhi (talk | contribs) m (IGEM:Peking/2007/Transformation moved to IGEM:Peking/2007/Count:Transformation: add group name:"Count") |
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Latest revision as of 22:40, 20 August 2007
- Hold competent cells (from -80℃ refrigerator) on ice.
- Gently mix ligation product (10-15 μL) with cells, then bath in ice for 30 min.
- (Precisely!) Bath in 42℃ water for 90 sec.
- Hold again in ice for 2 min.
- Bath in 37℃ for 45 min. For plasmid pKO3 (and other temperature-sensitive strains), bath in 30℃ for 1 hour.
- Separate cells on petri-dishes, and cultivate them in 37℃ for 12 hours (for pKO3, 24 hours are necessary).
