Team:Gaston Day School/17 June 2009

 High Efficiency E. coli Thaw tube of NEB 10-beta Comp. E. coli cells on ice until ice disappears. Gently mix and pipette 50mL into transformation tube on ice. Add plasma DNA and carefully mix Suspend the well 8E on Plate 2 in 15x TE Add 5x of Plasmid to E. Coli Place on ice for 30 mins Heat shock at 42C for 30 seconds Ice for 5 mins</dt> Pipette 950microliters of room temp SOC</dt> Place 37C for 60 mins and shake every 15 mins</dt> Warm plates to 37C</dt> Spin 5 mins</dt> Pipette out SOC</dt> Resuspend in 150microliters SOC</dt> Mix tubes</dt> Spread 50-100microliters onto plate and incubate overnight at 37-42C</dt> Plate 1 = 100microliters</dt> Plate 2 = 50microliters</dt>