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Lab July 24 2009
From 2009.igem.org
9:55 am - subcultured the overnight culture of DH5α at approximately 1/100 (1 mL culture into approximately 99-100 mL) and put both flasks into the shaker
12.50 pm - Prepared 10 mL of CM1 buffer and CM2 buffer: Preparation of Competent Cells Protocol
1.13 pm - culture at 0.517 OD, started procedure.
Altered Transformation Protocol
2.17 pm - Dilute pUC18 plasmid to 20 uL at 1/10: 2 uL plasmid in 18 uL TE.
Add 3 uL diluted plasmid to each of the three tubes of 200 uL competent cells (two new, one old), and mix gently.
Incubate on ice for 30 min.
2.47 pm - Heat shock the mixes at 42oC for exactly 2 min.
Transfer to ice for 1-2 min.
Add 400 uL of 37oC LB.
2.55 pm - Incubate for 1 hr 15 min at 37 oC in a shaking water bath.
4.10 pm - Plate 0.2 mL on Amp + LB plates.
Incubate plates at 37 oC approximately 18-24 hr.
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