phi29 DNA Polymerase (10 U/μL)
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phi29 DNA Polymerase (10 U/μL)
Thermo Scientific™

phi29 DNA Polymerase (10 U/μL)

Thermo Scientific phi29 DNA Polymerase is a highly processive polymerase featuring strong strand displacement activity, which allows for highly efficientRead more
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Catalog NumberQuantity
EP0091250 U
EP00921000 U
EP00945000 U
Catalog number EP0091
Price (USD)
94.50
Each
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Quantity:
250 U
Request bulk or custom format
Price (USD)
94.50
Each
Add to cart

Thermo Scientific phi29 DNA Polymerase is a highly processive polymerase featuring strong strand displacement activity, which allows for highly efficient isothermal DNA amplification. Produced from E. coli cells with a cloned gene 2 of Bacillus subtilis phage phi29, phi29 DNA Polymerase possesses 3'→5' proofreading exonuclease activity that acts preferentially on single-stranded DNA or RNA.

Features of phi29 DNA Polymerase
• Excellent processivity and strand displacement activity—more than 70 kb long DNA stretches can be synthesized
• Highly accurate DNA synthesis
• High yields of amplified DNA even from small amounts of template

Applications
Phi29 DNA Polymerase may be used in a wide variety of applications including:
• Rolling circle amplification (RCA)—generation of periodic DNA nanotemplates
• Multiple displacement amplification (MDA)
• Unbiased amplification of whole genome (WGA) (see figure below)
• Amplification of DNA for SNP and STR detection
• Cell-free amplification of DNA from single cells, pathogenic organisms, or metagenomes
• DNA template preparation for sequencing
• Protein-primed DNA amplification
• Recombination based-cloning
• Cell-free cloning of lethal DNA
• RNA-primed DNA amplification

Notes
1. The addition of pyrophosphatase to the reaction mixture with phi29 DNA Polymerase may enhance DNA synthesis.
2. Because of the enzymes’s 3’→5’ proofreading exonuclease, 3'-modified primers are highly recommended.

For Research Use Only. Not for use in diagnostic procedures.
Specifications
Product TypeStand-alone enzyme
For Use With (Application)MDA-WGA, RCA
Polymerasephi29 DNA Polymerase
Quantity250 U
Optimal Reaction Temperature30°C
Shipping ConditionDry Ice
Concentration10 U/μL
Reaction Time4 hours
Unit SizeEach
Contents & Storage
• phi29 DNA Polymerase (250 units at 10 U/μL)
• 10X Reaction Buffer for phi29 DNA Polymerase
• Store at –15 to –25°C.

Frequently asked questions (FAQs)

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

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

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

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

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.

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

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.

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

Can phi29 DNA Polymerase be heat-inactivated?

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

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

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