IGEM:UBC/2009/Notebook/UBC iGEM 2010/2010/06/08

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QS Track Transformed cell cultures from yesterday (Monday) were checked at 10:00 am today. Every agar plate had colonies growing. Next step: PCR the iGEM part to verify that the colonies do contain the parts. Competent Cell Creation See standard UBC iGEM protocol for detailed procedures. Overnight cultures of DB3.1 and DH5(alpha) were used to inoculate 2 * 50 mL cultures of each strain (11:00 am). The 50 mL cultures were grown at 30 °C. At 2:00 pm, each culture reached ~OD 0.400. We lost 1 DH5(alpha) flask due to contamination. Each 50 mL culture was split into 2 centrifuge tubes. One of the tubes was leaky, so 25 mL DB3.1 was lost. Other (no specific track) Tec-Monterrey iGEM Team requested E.Coli strain BW27783 which was used last year. Removed an aliquot of BW27783 competent cells from the -80 degree C freezer Kept the aliquot on ice Filled 2 14mL round bottom test tubes with 5mL of LB broth each Dipped a pipette tip into the now liquid aliquot of BW27783 cells (time elapsed in ice bucket - 30 minutes) Dipped the pipette tip into the broth of one of the tubes, and pipetted up and down very gently to mix ensure the cells were in the broth Placed both tubes (one with cells, one acting as a control) in a shaking incubator set to 30 degrees C at noon (12:00pm) Tube with cells was slightly cloudy by 5pm, control tube remained clear Left overnight