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12/07/13
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Revision as of 10:23, 14 August 2013 by AliceHaworth (Talk | contribs)
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Transformation of RFP control plasmid
- Keep competent cells on ice
- Add 50ul of cells to 4 tubes
- Add 1ul of resuspended DNA to 2 tubes
- Add 1ul RFP to the other 2 tubes
- Incubate for 30mins
- Heat shock at 42 degrees for 60secs
- Incubate on ice for 5mins
- Add 200ul of SOC media
- Incubate at 37degrees for 2hrs
- Spread on 6 plates:
- Positive control (normal plate)
- Negative control (No plasmid)
- 20ul of 50pg/ul
- 200ul of 50pg/ul
- 20ul of 10pg/ul
- 200ul of 10pg/ul
- leave plates on bench overnight
