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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

X-Gal staining protocol for beta galactosidase (blue/white screening)

(Estimated time: 20 min)

The principle is that X-Gal (5-bromo-4-chloro-3-indolyl-b-D-galactopyranoside) turns blue when reacts with beta-galactosidase. Mix 20 ul X-Gal 20 mg/ml and 80 ul SOC and spread on LB+amp plates (X-Gal and DMF are toxic, use face-mask for your safety!!! X-Gal is light-sensitive, remember to keep it in the dark, when possible) Incubate with 200 ul bacteria Result: blue colonies express LacZ, while white colonies don't.

40 mg/ml X-gal in DMF SOC medium Pre-warmed LB agar plate Face mask

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