Muratore:Protocols/Transformation
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< Muratore:Protocols(Difference between revisions)
Current revision (21:11, 8 June 2011) (view source) |
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##* spread remaining 800 μL on fresh plates if poor/no growth observed | ##* spread remaining 800 μL on fresh plates if poor/no growth observed | ||
##* otherwise, discard remainder after ~24h | ##* otherwise, discard remainder after ~24h | ||
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| + | [[Category:Protocol]] | ||
| + | [[Category:DNA]] | ||
| + | [[Category:E. coli]] | ||
Current revision
Bacterial Transformation
- 200 μL chemically- competent E. coli + 5 μL DNA
- 30 min on ice
- 30 sec @ 42°C
- 2 min on ice
- add 800 μL SOC
- 45 min @ 37°C
- spread 200 μL on rich-media plates
- → 37°C O/N (inverted)
- store remainder @ 4°C
- spread remaining 800 μL on fresh plates if poor/no growth observed
- otherwise, discard remainder after ~24h
