IGEM:UBC/2009/Notebook/UBC iGEM 2010/2010/08/30
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Biofilm TrackMelodyBiofilm Growth Protocol Day 4
Biofilm Growth Protocol Day 4
100823E Data
100823M Data
10823ERS Data
100823MRS
Biofilm Growth Protocol Day 1
QS
PCR Master mix Reagent 1x rxn volume (uL) Master Mix 5x rxn buffer 5 x15 75 10mM dNTP 0.5 x15 7.5 sdH2O 12.15 x15 182.25 Phusion polymerase 0.1 x15 1.5 MgCl2 2 x15 30 DMSO - 5% 1.25 x15 18.75 10uM fw primer (G1004) 2 x15 30 10uM re primer (G1005) 2 x15 30 Total 25 375
PCR Cycles: * 98C @ 3min * Cycle 27x: o 98C @ 10 sec o 72C @ 30 sec o 72C @ 40 sec o 72C @ 10 min * 10C @ hold * Ran gel for 1 hour. 80 V 0.5X TBE 1.3% agaorse. Two small 8-lane gel boxes. Also included 1 kb NEB ladders. * Results show many bands. Apparent weak bank/smudge of DNA above 2 kb region. Probably means many different plasmids present. The film probably has low antibiotic concentration. * Therefore, streaked some bacteria from each of the agrAC plates in the hopes of finding single colonies. Incubate at 37 °C. * Made DH5alpha O/N for competent cell making. Unfortunately, had to use Finlay lab incubator on the 3rd floor, which runs at 37 °C and 200 rpm.
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