Muratore:Protocols/Transformation

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(New page: ==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 μ...)
Current revision (20:11, 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]]
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[[Category:DNA]]
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[[Category:E. coli]]

Current revision

Bacterial Transformation

  1. 200 μL chemically- competent E. coli + 5 μL DNA
  2. 30 min on ice
  3. 30 sec @ 42°C
  4. 2 min on ice
  5. add 800 μL SOC
  6. 45 min @ 37°C
    1. spread 200 μL on rich-media plates
    2. → 37°C O/N (inverted)
    3. store remainder @ 4°C
      • spread remaining 800 μL on fresh plates if poor/no growth observed
      • otherwise, discard remainder after ~24h
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