phi29 HT DNA Polymerase

Content

500U

Features

• lStrong strand displacement capability

• lContinuous synthesis capability

• lHigh fidelity of amplification

Applications

• lDNA isothermal rolling loop amplification

• lDNA synthesis requiring strong strand displacement

• lRapid replication of plasmids

• lRapid replication requiring high fidelity

Advantages

The high temperature reaction characteristic of phi29 HT DNA Polymerase has the following advantages:

• lIn the next generation sequencing (NGS), the enzyme has stronger amplification activity for complex templates such as high GC content and palindrome structure, which makes the coverage of NGS more uniform and reduces the depth required for sequencing

• lThe production of whole genome amplification (WGA) products is increased for 2-5 times, and the amplification time for library construction is shortened to 1-2 hours.

• lThe gap region in sequencing is reduced, which can improve the quality and integrity of the data of single cell sequencing

• lThe results show that the quality and integrity of multiple displacement amplification (MDA) and rolling circle amplification (RCA) are improved, the non-specific amplification products are reduced, and the amplification performance and specificity of both MDA and RCA are improved.

Storage

Stored at -20°C for 3 years.

Notes

• l1 x phi29 Buffer: 50 mM Tris-HCl pH 7.5, 10 mM MgCl₂, 10 mM (NH₄) ₂SO₄, 4 mM DTT. If necessary, the final concentration of 1 mM DTT and 0.2 mg/ml BSA can be added separately to improve the reaction efficiency.

• lThe optimum reaction temperature of the enzyme is 30℃.

• lThe enzyme can be inactivated at 65℃ in 10 min.

• lAdjust the concentration of dNTP from 100 to 500 μM according to the type of experiments.

• lAddition of Yeast Pyrophosphatase to the reaction mixture with phi29 DNA Polymerase may enhance DNA synthesis.

• lThio-modification at the 3' end of the primers can avoid primer degradation.