dspB Track
- cPCR on plates (Streaked)
Colony PCR
PCR Master mix
Reagent | 1x rxn volume (uL) |
5x rxn buffer | 5 |
10mM dNTP | 0.5 |
sdH2O | 12.15 |
Phusion polymerase | 0.1 |
MgCl2 | 2 |
DMSO - 5% | 1.25 |
10uM fw primer (G1004) | 2 |
10uM re primer (G1005) | 2 |
Total | 25 |
PCR Tubes: His, no his, no his A, His A, W
- 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: 6:46pm
End: 7:49pm
Gel verification colony PCR products
- Protocol: gel verification protocol in Protocol (SOP)
- Changes: 1% agarose gel
- Machine conditions: 0.5x TBE buffer, 100V, 60min
Gel orientation:
Gel orientation
column 1: his | column 5:no-his | column 6: 100bp ladder | column 8: his A | column 9: no his A | column 10: W |
Results:
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