IGEM:UBC/2009/Notebook/UBC iGEM 2010/2010/07/12

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PCR genomic preps (PCR I)
Start from the beginning again with the A. pleurpneumoniae strains. PCR out dspB from the strains, dilute product, PCR dspB again, extract dspB from gel (purification).

Strains:
 * H49: 80ng/uL x2
 * H80: 80ng/uL x2
 * H171: 60ng/uL x2
 * Total: 6 samples + 1 water control + 2 extra = 9 samples

Protocol: Common protocols for PCR

2 Master Mixes:
 * MMA (master mix A): His-tagged FW primer - 6 samples + 1 H2O water + 2 extra = 9 samples
 * MMB (master mix B): FW primer - 6 samples + 2 extra = 8 samples
 * Total 17 samples in all for master mix (13 PCR tubes in total - 12 samples & 1 H2O control)

PCR Cycles:
 * 95C @ 2 min
 * Cycle 30x:
 * 95C @ 30 sec
 * 65C @ 10 sec
 * 72C @ 80 sec
 * 72C @ 10 min
 * 10C @ hold

Start: 1316 End: 1446

Gel Verification
Protocol: gel verification protocol in iGEM training manual Gel orientation: *Note: the second layer are Jason's samples - he is testing whether his PCR from last day worked or not

Machine conditions: 110V, 45 min, 0.5X TBE Buffer Results: Vicki Ma 23:00, 12 July 2010 (EDT)


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