From 2009.igem.org
Spin 1.5mL of overnight culture for 30 sec (5') in microfuge
Aspirate off all but 100x of the supernatant and resuspend the pellet by vortexing
Add 300x of TENS and mix by inversion. The solution should become viscous
Add 150x of sodium acetate and vortex. A fine white precipitate should form
Centrifuge for 2.5 min at 10k
TRANSFER the supernatant to a clean tube and add 2 volumes (1mL) of room temperature EtOH
Vortex and pellet DNA by centrifugation for 2-5 minutes at 10k
Wash pellet w/ 70% ethanol and allow the pellet to dry
Resuspend the pellet in 20x of TE w/ RNAseA
Digest 5-10x as usual
High eff. ecoRI: RFP, cRFP
Thaw e. Coli: till ice disappears
Pipette 100x into tube (x2)
Add 10x of Plasmid to E.coli
Place on ice for 30 min
Heat shock at 42C for 30 sec
Ice for 5 mins
Add 950x SOC
Place at 37C for 60 mins
Spin 5 mins
Pipette out SOC
Resuspend in 150x SOC
Spread 50x 8 100x amp onto new plates
8 tubes- grow overnight culture of Berkley plasmid
Make gel
Run gel electrophoresis
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