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

iGEM Project name 1

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dspB Track Transformants from yesterday taken out @ 1100 Transformant Results Plate # of colonies 1 6 colonies 2 no colonies 3 4 colony 4 6 colonies Protocol: Common protocols for PCR PCR Master mix Reagent 1x rxn volume (uL) Master Mix 10x rxn buffer 2.5 x18 45 10uM FW primer 2 x18 36 10uM RE primer 2 x18 36 10mM dNTP 2 x18 36 sdH2O 13.05 x18 234.9 Taq polymerase 0.2 x18 3.6 DMSO 1.25 x18 22.5 MgCl2 2 x18 36 Total 25 450 PCR Tubes From plate 1: 1T1 1T2 1T3 1T4 1T5 1T6 From plate 3: 3T1 3T2 3T3 3T4 From plate 4: 4T1 4T2 4T3 4T4 4T5 4T6 W (H2O control) PCR Cycles: 95C @ 10min Cycle 25x: 94C @ 30 sec 56C @ 30 sec 68C @ 2 min 68C @ 20 min 10C @ hold Start: 1204 End: 1351 Gel verification for colony PCR products Protocol: gel verification protocol in Protocol (SOP) Changes: 1% agarose gel Machine conditions: 0.5x TBE buffer, 100V, 50min Gel orientation: Gel orientation 1T1 1T2 1T3 1T4 1T5 1T6 3T1 3T2 3T3 100bp ladder 3T4 4T1 4T2 4T3 4T4 4T5 4T6 W (control) Results: Note: bottom results are QS Track's Vicki Ma 02:47, 11 August 2010 (EDT)