Team:Newcastle University/Protocols
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== Transformation after ligation == | == Transformation after ligation == |
Revision as of 14:13, 8 September 2008
Newcastle University
GOLD MEDAL WINNER 2008
Home | Team | Original Aims | Software | Modelling | Proof of Concept Brick | Wet Lab | Conclusions |
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Isolating Plasmid from Cells (Miniprep)
Restricting Plasmids (Double Restriction)
Making Frozen Glycerol Cell Stocks
Transformation after ligation
- Add 2μl plasmid to 50μl of competent cells.
- Incubate for 30 minutes on ice.
- Heat shock for 2 minutes at 42°C.
- Add 250μl LB.
- Incubate for 120 minutes at 37°C whilst shaking or rotating.
Making Frozen Glycerol Cell Stocks
- Pippette 2μl overnight culture into an Eppendorf tube.
- Add 0.5ml glycerol and vortex briefly to mix.
- Prepare a freeze box by decanting a small amount of liquid nitrogen into the box. (N.B: wear protective gloves and face mask.)
- Place the tube in the box and keep at -80°C.