17/07/13
From 2013.igem.org
(Difference between revisions)
AliceHaworth (Talk | contribs) (Created page with "<div style="font-family:arial;padding:5px;border-radius:5px;border:5px solid #FF2800;"> {| style="color:#87EA00;background-color:#FFFFFF;" cellpadding="2" cellspacing="2" border=...") |
AliceHaworth (Talk | contribs) (→Double digest) |
||
(3 intermediate revisions not shown) | |||
Line 12: | Line 12: | ||
|} | |} | ||
</div> | </div> | ||
+ | |||
+ | |||
+ | |||
+ | ==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 |
Latest revision as of 11:15, 14 August 2013
Home | Team | Official Team Profile | Project | Parts Submitted to the Registry | Modeling | Notebook | Safety | Attributions |
---|
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