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17/07/13
From 2013.igem.org
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*Use thermo scientific mini kit | *Use thermo scientific mini kit | ||
*Measure concentration using nanodrop | *Measure concentration using nanodrop | ||
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| + | ==Double digest== | ||
| + | |||
| + | *50ng of DNA | ||
| + | *2ul of 3.1 buffer | ||
| + | *1ul of each restriction (XbaI and PstIin the 6 samples, EcoRI and PstI in the 6 samples, 12 samples in total) | ||
| + | *Water to make total volume to 20ul | ||
| + | *Incubate at 60mins at 37 | ||
| + | *20mins at 80 | ||
Latest revision as of 11:15, 14 August 2013
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Isolating RFP plasmid
- Add 1.5ml of overnight culture to epindorph
- Centrifuge for 2mins
- Remove supernatent
- Repeat
- Use thermo scientific mini kit
- Measure concentration using nanodrop
Double digest
- 50ng of DNA
- 2ul of 3.1 buffer
- 1ul of each restriction (XbaI and PstIin the 6 samples, EcoRI and PstI in the 6 samples, 12 samples in total)
- Water to make total volume to 20ul
- Incubate at 60mins at 37
- 20mins at 80
