Glasgow/Wetlab/Protocols
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== Transforming BioBricks == | == Transforming BioBricks == | ||
- | # | + | #Split the competent ''E. coli'' cells (~75ul) in two. |
- | # | + | #Competent cells (~33ul) into eppendorfs on ice. |
- | # | + | #Add 1µl of plasmid DNA. |
- | # | + | #Incubate on ice for at least 5 mins. |
- | # | + | #Heat shock +42 ºC for 45 secs. |
- | # | + | #Back on ice for 2 mins to reduce damages. |
- | # | + | #Add 1ml of soc media. |
- | # | + | #Incubate +37ºC in a shaker for 1 hour. |
- | # | + | #Spread the reaction on kan/carb LB plates. |
- | # | + | #Grow +37 ºC overnight. |
Revision as of 17:26, 19 July 2007
Transforming BioBricks
- Split the competent E. coli cells (~75ul) in two.
- Competent cells (~33ul) into eppendorfs on ice.
- Add 1µl of plasmid DNA.
- Incubate on ice for at least 5 mins.
- Heat shock +42 ºC for 45 secs.
- Back on ice for 2 mins to reduce damages.
- Add 1ml of soc media.
- Incubate +37ºC in a shaker for 1 hour.
- Spread the reaction on kan/carb LB plates.
- Grow +37 ºC overnight.