IGEM:UBC/2009/Notebook/UBC iGEM 2010/2010/08/25

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dspB Track

 * O/N culture taken out at 1003
 * Transformants taken out at 1003
 * Results: 1,4,5,8,11,14,15,his,no-his: TMTC
 * Results: AA,TB: no colonies

Colony PCR
PCR Tubes: 1,4,5,8,11,14,15,his,n1,n2,n3,n4,n5,W(H2O control)
 * Pick colony from respective plate and place in respective tubes

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

Start: 1148 End: 1251

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

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

Restriction Digest Chlor backbone psb1C3

 * Protocol: biobrick (digesting)
 * Add: 5uL of backbone + 5uL of insert
 * Enzymes: EcoRI and PstI (1uL each to each tube)

Biofilm Growth Protocol Day 3 (100823E + M)

 * 100823E removed from incubator @ 1100
 * Controls showed no growth (woot woot)
 * Followed procedure and placed in biosafet cabinet overnight
 * 100823M removed from incubator @ 1335
 * Growth in control wells C4, C6, D10
 * Some showed a distinct colony
 * Contamination in growth wells C9, E5, D9
 * Black dots - looked like growth of some other bacteria

Biofilm Growth Protocol Day 2 (100824E + M)

 * Removed O/N cultures from incubator @ 1100 (20 hrs growth)
 * Plate layout below:

100824E + M Plate Layout

 * Note:
 * Controls chosen at random
 * R = RN4220
 * 8 = 8325-4
 * C = control
 * 100824M into incubator @ 1635
 * 100824E into incubator @ 1642
 * 100824E-E10 not fully filled, disregard all data from it


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