Team:Gaston Day School/9 July 2009

 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</dt> Resuspend the pellet in 20x of TE w/ RNAseA</dt> Digest 5-10x as usual</dt> High eff. ecoRI: RFP, cRFP</dt> Thaw e. Coli: till ice disappears</dt> Pipette 100x into tube (x2)</dt> Add 10x of Plasmid to E.coli</dt> Place on ice for 30 min</dt> Heat shock at 42C for 30 sec</dt> Ice for 5 mins</dt> Add 950x SOC</dt> Place at 37C for 60 mins</dt> Spin 5 mins</dt> Pipette out SOC</dt> Resuspend in 150x SOC</dt> Spread 50x 8 100x amp onto new plates</dt> 8 tubes- grow overnight culture of Berkley plasmid</dt> <dt>Make gel</dt> <dt>Run gel electrophoresis</dt>