ALLin™ Mega HiFi DNA/PFU Polymerase: High-fidelity, fast, and efficient PCR for complex templates.

Product Description

Derived from our HiFi polymerase, the highQu ALLin™ Mega HiFi DNA Polymerase provides much lower error rate PCR

with a 100 higher fidelity compared to Taq. The ALLin™ Mega HiFi DNA Polymerase is engineered to be much faster

and to generate a higher yield of long PCR products up to 20 kb from complex GC-rich templates.

Therefore the ALLin™ Mega HiFi DNA Polymerase is an excellent choice for longer and very complex PCR applications

where the highest fidelity is demanded. It is an enzyme of choice for cloning and all kind of sequencing applications including NGS.

Generated blunt-ended PCR products are suitable for ligation into blunt vectors. 

For maximum convenience, the ALLin™ Mega HiFi Mastermix, 2X (HLM0201) and ALLin™ Mega HiFi Red Mastermix,

2X (HLM0301) are available. Pfu DNA Polymerase was originally found in thermophilic bacterium Pyrococcus furiosus. It is a thermostable Pfu polymerase that was widely used in molecular biology applications for its high fidelity of DNA amplification due to a high proofreading activity. Pfu polymerase has 3' to 5' exonuclease proofreading activity, which enables it to correct errors that occur during DNA synthesis.

The enzyme is commonly used for cloning and sequencing applications, however it is relatively slow, sensitive to contaminants

and inhibitors and has low processivity resulting in usually low  PCR  yields.

Alternative, mostly better  DNA polymerases  have been developed by different companies to address the limitations of Pfu polymerase.

These include such enzymes as Phusion® DNA polymerase, Q5® DNA polymerase from NEB, and many others.

These enzymes offer improved elongation rates, reduced error rates, and increased efficiency for NGS applications.

The ALLin™ MEGA HiFi Pfu Polymerase offered by highQu provides 100X higher fidelity compared to Taq Polymerase,

which is much higher than Pfu Polymerase. In addition, the hot start enzyme version allows to reduce the background to the minimum.

Such robust enzyme offers not only precise amplification, but also enables PCR from very complex templates

and generation of very long PCR products up to 30 kb and above.

Availability of faster, more robust and more precise polymerases influenced successful development

of next generation sequencing techniques (NGS) where PCR precision and yield is crucial for reliable sequencing results.

Applications

Fast high-fidelity PCR (up to 100 x Taq).

Long PCR up to 20 kb.

Amplification of complex (GC/AT-rich) templates.

Sequencing, including NGS.

Blunt-end cloning and other applications.

Benefits

Fast, high yield PCR with the fidelity 100x higher than Taq.

Up to 20 kb long PCR even from complex templates.

Increased processivity for faster amplification and higher yield.

High thermostability for better denaturation of GC rich templates.

Best choice for NGS and other sequencing applications.