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

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dspB Track

  • Transformants from yesterday taken out @ 1100
Transformant Results
Plate # of colonies
16 colonies
2no colonies
34 colony
46 colonies

Protocol: Common protocols for PCR

PCR Master mix
Reagent1x rxn volume (uL)Master Mix
10x rxn buffer2.5x1845
10uM FW primer2x1836
10uM RE primer2x1836
10mM dNTP2x1836
sdH2O13.05x18234.9
Taq polymerase0.2x183.6
DMSO1.25x1822.5
MgCl22x1836
Total25450
PCR Tubes
From plate 1:1T11T21T31T41T51T6
From plate 3:3T13T23T33T4
From plate 4:4T14T24T34T44T54T6
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
1T11T21T31T41T51T63T13T23T3100bp ladder3T44T14T24T34T44T54T6W (control)

Results:
File:100806 DspB 12.tif

  • Note: bottom results are QS Track's

Vicki Ma 02:47, 11 August 2010 (EDT)