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菌落PCR |
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Despite the advantages of screening transformants directly from cultures or plates, instead of first having to isolate DNA, the utility of colony PCR remains limited due to the inherent limitations of Taq polymerase in crude sample PCR. KAPA2G Robust DNA polymerase is a highly versatile 2nd generation enzyme that exhibits superior tolerance to a wide range of common PCR inhibitors and unrivalled performance in colony PCR.
Download the Colony PCR Application Note from Kapa Biosystems now to find out how to:
- PCR direct from cultures
- Eliminate dilutions of cultures
- Achieve up to 100% success working in E. coli and Yeast
KAPA2G Robust Exhibits Consistent Amplification Direct from Colonies and Overnight Cultures
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Amplification of a 2.7 kb insert from four commonly used E. coli strains (DH5a, DH10B, JM109 or BL21) using KAPA2G Robust HotStart DNA Polymerase (top) or a "robust" wild-type Taq (bottom). Colonies (grown on LB-agar + Amp plates) were either suspended directly in individual PCR reactions (left) or first resuspended in PCR grade water and then transferred to PCR tubes containing reaction master mixes (middle). For the right hand panel, overnight cultures (prepared in LB + Amp) were used directly in PCRs. |
 Amplification of a 2.5 kb (left) or 1.6 kb (right) fragmant of the GSH1 gene from three commonly used S. cerevisae strains (BY4742, FY23 and W303) using KAPA2G Robust HotStart DNA Polymerase (top) or a "robust" wild-type Taq (bottom). Colonies (from YPD-agar plates) or YPD overnight cultures were lysed in 50 ul volumes with NaOH or ZymolyaseTM(as indicated). Of each lysate, 2.5 ul wasused in the PCR. |
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