IGEM:UBC/2009/Notebook/UBC iGEM 2010/2010/09/24

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Biofilm Protocol Day 1

 * Note: Each "batch" of O/N cultures include one test tube of control, RN4220, and 8325-4
 * Note: I used the timer/clock on the lab bench for the timing


 * Batch 1: in @ 4:03 p.m.
 * Batch 2: in @ 5:10 p.m.
 * Batch 3: in @ 6:06 p.m.
 * Batch 4: in @ 4:03 p.m.
 * Batch 5: in @ 4:03 p.m.
 * Batch 6: in @ 4:03 p.m.

dspB Track

 * 1/100 dilution of pre-dspB and Pcon in 37C shaking incubator at 12:10pm
 * O/N culture in 4C


 * 1/100 dilution of Pcon growing in 37C shaking (AMP not CHLOR) at 3:30pm
 * 1/100 dilution of:
 * pre-dspB OD600=1.054
 * P-con OD600=1.344

PCR using Rafael's method Attempt #5

 * Using new PNKed primers and Taq


 * Tubes: rxn2, W

PCR Cycles:
 * 95C @ 2min
 * Cycle 30x:
 * 95C @ 30 sec
 * 58.5C @ 10 sec
 * 72C @ 80 sec
 * 72C @ 10 min
 * 10C @ hold

Start: 4:17pm End: 5:48pm

Gel verification colony PCR products
Gel orientation: Results:
 * Protocol: gel verification protocol in Protocol (SOP)
 * Machine conditions: 0.5x TBE buffer, 100V, 45min

cPCR NCC colonies to verify before making glycerol stock

 * Tubes: NCC, Poco, Pcon, W
 * Poco and Pcon are for QS track

PCR Cycles:
 * 98C @ 3min
 * Cycle 27x:
 * 98C @ 10 sec
 * 72C @ 30 sec
 * 72C @ 40 sec
 * 72C @ 10 min
 * 10C @ hold

Start: 7:26pm End: 8:29pm

Gel verification colony PCR products
Gel orientation: Results:
 * Protocol: gel verification protocol in Protocol (SOP)
 * Machine conditions: 0.5x TBE buffer, 100V, 60min

O/N culture

 * O/N culture of NCC made with 1.5mL LB broth for glycerol stock
 * used 1.5uL of C1 antibiotic
 * put into 37C shaking incubator at 6:40pm


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