phi29 DNA Polymerase (10 U/μL), 250 units - FAQs

View additional product information for phi29 DNA Polymerase (10 U/μL) - FAQs (EP0092, EP0094, EP0091)

8 product FAQs found

What is the error rate of phi29 DNA Polymerase?

The error rate of phi29 DNA Polymerase is 1 x 10-5.
The error rate of phi29 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

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Can I use phi29 DNA Polymerase to incorporate 5-methyl-dCTP?

phi29 DNA Polymerase should be able to incorporate 5-methyl-dCTP nucleotides and other modified nucleotides.
The incorporation of modified nucleotides by phi29 DNA Polymerase can be reviewed in the literature:
Jakubovska, J., Tauraitė, D., Birštonas, L., & Meškys, R. (2018). N 4-acyl-2′-deoxycytidine-5′-triphosphates for the enzymatic synthesis of modified DNA. Nucleic Acids Research, 46(12), 5911–5923. https://doi.org/10.1093/nar/gky435

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What is the minimal recommended time for amplification with phi29 DNA Polymerase?

The optimal reaction time for DNA amplification with phi29 DNA Polymerase is 4 hours. For samples with ≥1 pg of DNA input, DNA amplification time can be shortened to 2 hours if maximizing product yield is not essential.

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Can phi29 DNA Polymerase be used with liquid media culture or colonies?

Yes. phi29 DNA Polymerase can work with different types of sample input such as purified DNA, liquid media culture, agar plate colonies, etc.

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Can phi29 DNA Polymerase be heat-inactivated?

Yes, we recommend heat-inactivating phi29 DNA Polymerase at 65 degrees C for 10 min.

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Is phi29 DNA Polymerase supplied with dNTPs?

No. dNTPs can be purchased separately (Cat. No. R0181).

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Is phi29/EquiPhi29 DNA Polymerase suitable for amplification of short DNA fragments?

The amplification efficiency of MDA (multiple displacement amplification) reaction rapidly diminishes as the molecular weight of the starting material decreases, thus making the polymerase unsuitable for amplification of low-molecular weight DNA. However, it is suitable for amplification from extremely low amounts of starting DNA material, making it ideal for single cell analysis.

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What type of DNA template can be amplified by phi29 DNA Polymerase?

Both linear and circular DNA templates can be amplified by phi29 DNA Polymerase.

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