AQ97 High Fidelity DNA Polymerase 2x Master Mix is a convenient alternative to AQ97 High Fidelity DNA Polymerase, displaying the following features; high fidelity is measured to >60x Taq DNA Polymerase, fast amplification due to its high extension rate, ability to amplify problematic DNA targets, such as those with low to high GC content and ability to perform amplification on long DNA targets. These features enables accurate and reliable PCR results.
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
All-in-one 2x master mix for great convenience
Timesaving reaction set-up
High fidelity: >60x Taq Fidelity
3’→5’ proofreading exonuclease activity
High elongation rate: 10 sec/kb
Long-range capability: 11 kb gDNA
Good coverage on DNA templates with low to high GC content
DESCRIPTION
AQ97 High Fidelity DNA Polymerase 2x Master Mix is a ready to use 2x reaction 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 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.
FIDELITY
The fidelity of a polymerase refers to its ability to insert the correct base during PCR. On the contrary, the rate of misincorporation is known as a polymerases error rate. Fidelity depends on the polymerase, the buffer system in use and the quality of your template DNA.
FIDELITY: The fidelity values of AQ97 High Fidelity DNA Polymerase, AccuPol DNA Polymerase, high fidelity DNA polymerase P and high fidelity DNA polymerase Q was compared to the fidelity value of Taq DNA polymerase (1x) ERROR RATES: a Errors per base per doubling. Standard deviations are given in (). The presented error rates may not be comparable to those presented elsewhere due to technical and methodical differences. b Error rates for AQ97 High fidelity DNA Polymerase and also for the two high-fidelity DNA polymerases P and Q were below the detection limit for the method used. This limit is estimated to be 8.4 x 10-6 errors per base per doubling (60 x Taq fidelity). Click to enlarge picture.
LOW NUMBER OF SUBSTITUTIONS
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.
