Glasgow/Wetlab/Protocols

From 2007.igem.org

(Difference between revisions)
(Transforming BioBricks)
(Transforming BioBricks)
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== Transforming BioBricks ==
== Transforming BioBricks ==
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#split the competent ''E. coli'' cells (~75ul) in two.
+
#Split the competent ''E. coli'' cells (~75ul) in two.
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#competent cells (~33ul) into eppendorfs on ice..
+
#Competent cells (~33ul) into eppendorfs on ice.
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#add 1ul of plasmid DNA.
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#Add 1µl of plasmid DNA.
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#incubate on ice for at least 5 mins.
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#Incubate on ice for at least 5 mins.
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#heat shock +42 ºC for 45 secs.
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#Heat shock +42 ºC for 45 secs.
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#back on ice for 2 mins to reduce damages.
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#Back on ice for 2 mins to reduce damages.
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#add 1ml of soc media.
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#Add 1ml of soc media.
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#incubate +37ºC in a shaker for 1 hour.
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#Incubate +37ºC in a shaker for 1 hour.
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#spread the reaction on kan/carb LB plates.
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#Spread the reaction on kan/carb LB plates.
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#grow +37 ºC overnight.
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#Grow +37 ºC overnight.

Revision as of 17:26, 19 July 2007

Transforming BioBricks

  1. Split the competent E. coli cells (~75ul) in two.
  2. Competent cells (~33ul) into eppendorfs on ice.
  3. Add 1µl of plasmid DNA.
  4. Incubate on ice for at least 5 mins.
  5. Heat shock +42 ºC for 45 secs.
  6. Back on ice for 2 mins to reduce damages.
  7. Add 1ml of soc media.
  8. Incubate +37ºC in a shaker for 1 hour.
  9. Spread the reaction on kan/carb LB plates.
  10. Grow +37 ºC overnight.