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From 2009.igem.org

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	LB medium DNA resuspension from iGEM 2009 plates Transformation X-Gal staining protocol for beta galactosidase (blue/white screening) Glycerol stocks Plasmid extraction BioBrick digestion with restriction enzymes Electrophoresis DNA gel extraction DNA precipitation with sodium acetate Ligation PCR Sensing ethanol concentration: our do-it-yourself kit

DNA precipitation with sodium acetate

(estimated time: 1 hour)

We use it only if the miniprep did not give us a high yield of DNA Add 1/10 digestion volume of sodium acetate 3 M Add 2.5 digestion volume of absolute ethanol Freeze at -80°C for 30 min Centrifuge at 13000 rpm, 4°C for 20 min Decant supernatant Add 50 µl of 70% ethanol Centrifuge at 13000 rpm, 4°C for 20 min Remove all supernatant with a pipette Air dry pellet until ethanol is totally removed Elute with 5 µl ddH2O

Sodium acetate Absolute ethanol Ethanol 70% ddH2O Previously cut plasmid

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