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Newcastle University

GOLD MEDAL WINNER 2008

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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.

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 [[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===