Template:SSB-Protocols Col PCR: Difference between revisions
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| 10 mM dNTPs||1||1 | | 10 mM dNTPs||1||1 | ||
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| 10x buffer | | 10x Taq buffer ||5||5 | ||
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| oligo 1||1||1 | | oligo 1||1||1 | ||
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| template||1||1 | | template||1||1 | ||
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| Taq polymerase | | Taq polymerase ||0.25||0.25 | ||
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Latest revision as of 16:59, 20 June 2010
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 |
