Template:SSB-Protocols Col PCR
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Be sure to save some colonies on a block!
Do 4 colonies per transformation.
Pick colony and dab into PCR then dab into block.
Put block in incubator at 37C for 5-6hrs.
Use 33-50ul of PCR mix per colony.
Use PCR tubes!
Run colony PCR program based on fragment size.
Colony PCR Recipe:
H2O | 41.5 | 41.5 |
10 mM dNTPs | 1 | 1 |
10x Taq buffer | 5 | 5 |
oligo 1 | 1 | 1 |
oligo 2 | 1 | 1 |
template | 1 | 1 |
Taq polymerase | 0.25 | 0.25 |