Transformation

Date:

Ligation numbers:

Ligation date:

Cells:

Plates:

amp:

X-gal:

IPTG:

Add 2 æl ligation reaction to 50 æl competent cells.

Incubate 30 minutes on ice.

Heat shock 42ø C 45 seconds.

Incubate 2 minutes on ice.

Add SOC and incubate 60 minutes at 37ø C while shaking.

Plate aliquots (50 and 100 ul) of cells, incubate O/N 37øC in incubator.

Plates are LB with 50 æg/ml amp.

FYI: Plates spread with 4 æl IPTG 200 mg/ml (Laura Hall's @1M or 238 mg/ml) and 40 æl X-gal 20 mg/ml (2%). SOC is 2 g bactotryptone, 0.5 g yeast extract, 1 ml 1M NaCl, and 0.25 ml 1 M KCl in 97 ml H₂O, supplemented to 20 mM Mg++ with 2 M Mg++ stock (1 M MgCl₂ plus 1 M MgSO₄). Add 1 ml 2 M glucose to make media 20 mM. Filter.

Transformation scoring:

lig#/date	trans#/date	# colonies	white/blue	miniprep	master
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