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	== Transforming BioBricks ==		== Transforming BioBricks ==
-	#9 reactions including 5 tubes of competent cells (4 x Top 10, 1 x DB3.1).	+	#split the competent ''E. coli'' cells (~75ul) in two.
-	#split the content (~75ul) in two.	+
	#competent cells (~33ul) into eppendorfs on ice..		#competent cells (~33ul) into eppendorfs on ice..
	#add 1ul of plasmid DNA.		#add 1ul of plasmid DNA.

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Revision as of 17:24, 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 1ul 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.