AQ97 proofreading MasterMix 2X

AQ97 High Fidelity DNA Polymerase MasterMix 2X is a ready-to-use 2x PCR mix composed of AQ97 High Fidelity DNA Polymerase and an optimised buffer system including dNTPs and magnesium chloride, allowing robust amplification on DNA target with low to high GC content and long DNA targets. The DNA binding domain of this polymerase, ensures excellent high fidelity, long range capacity and also fast amplification results. AQ97 High Fidelity DNA Polymerase exhibits both 5'→3' DNA polymerase activity and 3'→5' proofreading exonuclease activity enabling this polymerase to correct base pair mismatches.

For longer targets than 11 kb we recommend to use our AQ97 High Fidelity DNA Polymerase with a capability of amplifying DNA targets up to 18 kb.. For difficult amplicons, such as GC-rich DNA samples, those with complex secondary structures or long amplicons, the addition of 1 – 2 M Betaine Enhancer Solution is recommended.

AQ97 High Fidelity DNA Polymerase is a novel proofreading DNA polymerase. AQ97 High Fidelity DNA Polymerase is a fused protein complex of DNA polymerase with a processivity-enhancing DNA binding domain. Alongside very fast and robust amplification of complex and long targets, AQ97 High Fidelity DNA Polymerase displays a high fidelity ensuring accurate amplification. The enzyme is well suited for PCR experiments that require amplification with very low error rates, such as cloning/sub-cloning, NGS applications, SNP analysis and mutagenesis.

The AQ97 master mix performs robust amplification of a vast range of difficult targets, including those up to 11 kb and with high to low GC content. The fidelity of the polymerase is more than 60x Taq DNA Polymerase. AQ97 High Fidelity DNA Polymerase 2X Master Mix is recommended for applications, which require extremely high fidelity such as cloning/Sub-cloning, NGS applications, SNP analysis and mutagenesis.

Features:
• High Fidelity: > 60x Taq fidelity
• High elongation rate: 10 sec/kb
• Long range amplification: 11 kb for human gDNA
• 3’ to 5’ proofreading exonuclease activity

Comparison figures of fidelity values for AQ97 DNA Polymerase, AccuPol DNA Polymerase, two well-recognized high fidelity DNA polymerases P and Q and Taq DNA Polymerase were determined through NGS-based analysis of nucleotide misincorporation during PCR. Initially, PCR amplification was performed on a ~ 200 bp synthetic DNA target, generating PCR products for each of the tested polymerases (using recommended setup conditions).

Distribution of substitution errors. PCR was performed using Taq DNA Polymerase, AQ97 High Fidelity DNA Polymerase, high fidelity DNA Polymerase Q and high fidelity DNA Polymerase P. The PCR was followed by NGS sequencing of the PCR products. The number of substitutions at each PCR target position was calculated and plotted in diagram A. Substitutions include misincorporated nucleotides and deletions at each position. Non-polymerase errors are subtracted from the total number of errors to revel true polymerase errors. Non-polymerase errors include mutations caused by thermocycling-induced DNA damage, pre-NGS sample preparation and sequencing errors. In these diagrams the average number of substitutions for Taq DNA Polymerase (Taq average) and for AQ97 High Fidelity DNA Polymerase (AQ97 average) is also plotted. Diagram B magnifies the area near the detection limit, displaying more information about the number of substitutions for AQ97 High Fidelity DNA Polymerase, high fidelity DNA Polymerase Q and high fidelity DNA polymerase P. Click to enlarge diagrams.