EquiPhi29™ DNA Amplification Kit, 100 Reactions - FAQs

Q: What is the error rate of EquiPhi29 DNA Polymerase?

The error rate of EquiPhi29 DNA Polymerase is 6 x 10 -6 . The error rate of EquiPhi29 DNA Polymerase was measured according to the method described in literature: Mielinis, P., Sukackaitė, R., Serapinaitė, A., Samoilovas, F., Alzbutas, G., Matjošaitis, K., & Lubys, A. (2021). MUA-based molecular indexing for rare mutation detection by Next-Generation sequencing. Journal of Molecular Biology, 433(19), 167209. https://doi.org/10.1016/j.jmb.2021.167209 Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center .

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5 个常见问题解答

What is the error rate of EquiPhi29 DNA Polymerase?

The error rate of EquiPhi29 DNA Polymerase is 6 x 10^-6.
The error rate of EquiPhi29 DNA Polymerase was measured according to the method described in literature:
Mielinis, P., Sukackaitė, R., Serapinaitė, A., Samoilovas, F., Alzbutas, G., Matjošaitis, K., & Lubys, A. (2021). MUA-based molecular indexing for rare mutation detection by Next-Generation sequencing. Journal of Molecular Biology, 433(19), 167209. https://doi.org/10.1016/j.jmb.2021.167209

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.

Can I use EquiPhi29 DNA Polymerase to incorporate 5-methyl-dCTP?

Thermo Scientific EquiPhi29 DNA Polymerase is a proprietary mutant phi29 DNA Polymerase developed through in vitro protein evolution. EquiPhi29 DNA Polymerase as well as phi29 DNA Polymerase should be able to incorporate 5-methyl-dCTP nucleotides and other modified nucleotides.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.

When using EquiPhi29 DNA Polymerase, do I need to purify amplified DNA products before downstream applications?

Cleaning of the amplified product is not required prior to several downstream methods (e.g., debranching, digestion with restriction endonucleases, Sanger sequencing); the dilution of amplified product is sufficient. If the clean-up procedure is needed, we recommend using an affinity-based spin-column or magnetic bead-based purification method.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.

What is the minimal recommended time for amplification with EquiPhi29 DNA Polymerase?

The optimal reaction time for DNA amplification with EquiPhi29 DNA Polymerase is 2 hours. For samples with ≥1 pg of DNA input, DNA amplification time can be shortened to 1 hour if maximizing product yield is not essential.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.

Can liquid media culture or colonies be used as a starting material for amplification with EquiPhi29 DNA Polymerase?

Yes. EquiPhi29 DNA Polymerase can work with different types of sample input material such as purified DNA, liquid media culture, agar plate colonies, etc.

Find additional tips, troubleshooting help, and resources within our PCR and cDNA Synthesis Support Center.