AQ97 Hot Start High Fidelity PCR 2x Master Mix

AQ97 Hot Start High Fidelity 2x Master Mix contains a proofreading enzyme for  robust amplification of DNA targets with low to high GC content and long DNA targets  of 18 kb. 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.

We recommend to use our AQ97 High Fidelity 2x Master Mix 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 master mix 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.

Features:

• High Fidelity: > 60x Taq fidelity
• High elongation rate: 10 sec/kb (up to 6000bp per minute)
• Built in hot start technology
• Long range amplification: 18 kb for human gDNA
• 5'-3' polymerase acitivity and 3'-5' exonuclease activity.
• Generates blund ends.

Picture 1: Comparison figures of fidelity values for AQ97 DNA Polymerase, AccuPol DNA Polymerase, and 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). 

Picture 2: Eight different human genomic DNA targets, ranging from 400 to 800 bp in length and with GC content between 29% and 78%, were amplified with AQ97 Hot Start High Fidelity DNA Polymerase 2x Master Mix. Robust amplification was observed for all targets. For targets with GC content greater than 70%, 2 M Betaine Enhancer Solution was included in the reaction mix.

Picture 3: Five different targets of human genomic DNA ranging from 2 kb to 17.5 kb were amplified. Robust amplification was observed for all targets using AQ97 Hot Start High Fidelity DNA Polymerase 2x Master Mix, demonstrating the ability of the master mix to amplify large and complex targets. Amplicon sizes are indicated at the top of the gel. Marker M: High Range DNA Ladder.