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JUNE: WEEK 5
June, 28thFirst time in wet lab 11 LB agar plates, 35 LB agar + Amp plates and 15 LB agar + Cm 12.5 were prepared.
For each of them 1 μl was transformed in 100 μl of home-made TOP10 competent cells. Transformants were plated on LB agar plates with the right antibiotic and incubated ON at 37°C. June, 29thIn the morning all plates showed colonies (BBa_J04450 showed light red colonies); for each we picked one colony and inoculated it in 1 ml of liquid LB with the proper antibiotic (ampicillin for BBa_C0060, BBa_C0061 and BBa_K081022, kanamycin for BBa_J04450). Cultures were grown at 37°C, 220 rpm. In the afternoon glycerol stocks were prepared (750 μl of cultures and 250 μl of 80% glycerol) and stored at -80°C except for BBa_C0061 which was not sufficiently grown (we let it grow until next morning); then cultures were refilled with 5 ml of liquid LB with the proper antibiotic and incubated ON at 37°C, 220 rpm. June, 30thAll cultures were in saturation growth phase; glycerol stock for BBa_C0061 Plasmids were extracted with MiniPrep kit; purified DNA was quantified with NanoDrop Spectrophotometer:
BBa_B0030, BBa_B0031, BBa_B0032, BBa_B0034, BBa_B0015, pSB4C5, BBa_I13501, BBa_I13507 were inoculated from freezer glycerol stocks and grown ON at 37°C, 220 rpm for MiniPrep.
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Team:UNIPV-Pavia/Calendar/June/week5
From 2011.igem.org
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Latest revision as of 09:50, 18 September 2011