IGEM:UBC/2009/Notebook/UBC iGEM 2010/2010/08/31

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Melody

 * Took out O/N cultures of RN4220 and 8325-4 from non-shaking incubator @ 11:52 a.m.

Biofilm Protocol Day 2

 * Innoculated plates 100830M120+100830M222

100830M1 + 100830M2 96-well plate Layout

 * Note:
 * Controls chosen at random
 * R = RN4220
 * 8 = 8325-4
 * C = control


 * Incubated @ 2:00 p.m. w/o shaking

Overnight cultures

 * 2 O/N cultures of RN4220 and 8325-4
 * one in incubator w/ shaking
 * one in incubator w/o shaking


 * Both in @ 3:57 p.m.

DspB Track
PCR off gDNA for more PCR products


 * To His1,His2,His3,His4: add 2uL of up-his & dw primers each
 * To no his1,no his2,no his3,no his4: add 2uL of up & dw primers each
 * To all, add 5uL of DNA

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

Start: 1234 End: 1335

Gel verification colony PCR products
Gel orientation: Results:
 * Protocol: gel verification protocol in Protocol (SOP)
 * Changes: 1% agarose gel
 * Machine conditions: 0.5x TBE buffer, 100V, 60min

PCR purification

 * All 4 tubes of His in 1 spin column (88uL + 440uL PBI)
 * All 4 tubes of no his in 1 spin column (88uL + 440uL PBI)
 * [His] = 126.3ng/uL
 * [No his] = 196.8ng/uL