Glasgow/Wetlab/Protocols

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== Transforming BioBricks (Maija)
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== Transforming BioBricks ==
#9 reactions including 5 tubes of competent cells (4 x Top 10, 1 x DB3.1).
#9 reactions including 5 tubes of competent cells (4 x Top 10, 1 x DB3.1).
#split the content (~75ul) in two.
#split the content (~75ul) in two.

Revision as of 17:24, 19 July 2007

Transforming BioBricks

  1. 9 reactions including 5 tubes of competent cells (4 x Top 10, 1 x DB3.1).
  2. split the content (~75ul) in two.
  3. competent cells (~33ul) into eppendorfs on ice..
  4. add 1ul of plasmid DNA.
  5. incubate on ice for at least 5 mins.
  6. heat shock +42 ºC for 45 secs.
  7. back on ice for 2 mins to reduce damages.
  8. add 1ml of soc media.
  9. incubate +37ºC in a shaker for 1 hour.
  10. spread the reaction on kan/carb LB plates.
  11. grow +37 ºC overnight.