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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 | ||
Latest revision as of 10:23, 14 August 2013
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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
