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Revision as of 17:26, 19 July 2007 (view source) MaijaP (Talk | contribs) (→Transforming BioBricks) ← Older edit		Revision as of 17:27, 19 July 2007 (view source) MaijaP (Talk | contribs) (→Transforming BioBricks) Newer edit →
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	#Incubate on ice for at least 5 mins.		#Incubate on ice for at least 5 mins.
	#Heat shock +42 ºC for 45 secs.		#Heat shock +42 ºC for 45 secs.
-	#Back on ice for 2 mins to reduce damages.	+	#Back on ice for 2 mins to reduce damage.
	#Add 1ml of soc media.		#Add 1ml of soc media.
	#Incubate +37ºC in a shaker for 1 hour.		#Incubate +37ºC in a shaker for 1 hour.
	#Spread the reaction on kan/carb LB plates.		#Spread the reaction on kan/carb LB plates.
	#Grow +37 ºC overnight.		#Grow +37 ºC overnight.

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Revision as of 17:27, 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 damage.
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.