Colony PCR
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Colony PCR is can be used after a transformation to screen colonies for the desired plasmid. Primers are used which generate a PCR product of known size. Thus, any colonies which give rise to an amplification product of the expected size are likely to contain the correct DNA sequence. | Colony PCR is can be used after a transformation to screen colonies for the desired plasmid. Primers are used which generate a PCR product of known size. Thus, any colonies which give rise to an amplification product of the expected size are likely to contain the correct DNA sequence. | ||
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==Specific Protocols== | ==Specific Protocols== | ||
*[[Endy:Colony PCR]] | *[[Endy:Colony PCR]] | ||
| - | *[[Knight:Colony PCR]] | + | *[[Knight:Colony PCR]] |
| + | *[[McClean:_Colony_PCR_(Yeast)]] | ||
| + | *[[Silver:_Colony_PCR]] | ||
| + | *[[Blackburn:Yeast_Colony_PCR]] | ||
==Notes== | ==Notes== | ||
#I have compared Taq and Vent using this reaction mix and found that Taq worked better. For very long amplicons, I would recommend using Phusion, which has its own reaction mix. I have not found a difference between Phusion and Taq on amplicons shorter than ~3kb. | #I have compared Taq and Vent using this reaction mix and found that Taq worked better. For very long amplicons, I would recommend using Phusion, which has its own reaction mix. I have not found a difference between Phusion and Taq on amplicons shorter than ~3kb. | ||
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| + | [[Category:Protocol]] | ||
| + | [[Category:Escherichia coli]] | ||
| + | [[Category:DNA]] | ||
Current revision
| back to protocols | ||
General Information
Colony PCR is can be used after a transformation to screen colonies for the desired plasmid. Primers are used which generate a PCR product of known size. Thus, any colonies which give rise to an amplification product of the expected size are likely to contain the correct DNA sequence.
Specific Protocols
- Endy:Colony PCR
- Knight:Colony PCR
- McClean:_Colony_PCR_(Yeast)
- Silver:_Colony_PCR
- Blackburn:Yeast_Colony_PCR
Notes
- I have compared Taq and Vent using this reaction mix and found that Taq worked better. For very long amplicons, I would recommend using Phusion, which has its own reaction mix. I have not found a difference between Phusion and Taq on amplicons shorter than ~3kb.
