AQ90 High Fidelity DNA Polymerase is a proofreading DNA polymerase displaying the following features; high fidelity is measured up to 50x Taq DNA Polymerase, 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 feature enables accurate and reliable PCR results.
AQ90 High Fidelity DNA Polymerase is recommended for applications, which require extremely high fidelity such as cloning/sub-cloning, NGS applications and mutagenesis.
High fidelity – measured up to 50x Taq Fidelity
3’→5’ proofreading exonuclease activity
Exceptional low error rate
Processes up to 8.5 kb gDNA and ≤ 12.5 kb for λDNA
Good coverage on DNA templates with low to high GC content
AQ90 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.
AQ90 High Fidelity DNA Polymerase kit is supported with 10x AQ90 Buffer allowing robust amplification on DNA targets with low to high GC content and long DNA targets. The kit also includes an extra tube of MgCl2 in 25 mM concentration if adjustment of PCR conditions is required.
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.
For more convenient handling AQ90 High Fidelity DNA Polymerase is also available as a 2x master mix
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.
Performance of AQ90 High Fidelity DNA Polymerase was compared to three leading High Fidelity DNA Polymerases (A, B and C). Nine different human genomic DNA targets, 400 – 800 bp in length and with GC content ranging from 29 – 78 %, were amplified. Robust amplification was observed using AQ90 High Fidelity DNA Polymerase on the nine human genomic targets. In contrary, none of the three leading High Fidelity DNA polymerase were able to provide the same level of robust amplification on the DNA targets with the higher GC content, under the conditions tested here. Amplification studies has been set up, as recommended by the manufactures. Tm Calculators of the respective competitors were used to calculate optimal annealing temperature for primers. When amplifying GC-rich targets, 2 M Betaine Enhancer Solution (AQ90 DNA Polymerase), GC enhancer (Competitor A and B) or GC-rich specific PCR Buffer (competitor C) were included in the reaction mix.
LONG RANGE AMPLIFICATION
AQ90 High Fidelity DNA Polymerase enables amplification of large and complex amplicons. Four different targets of human genomic DNA ranging from 2 kb and up to 8.5 kb was used in this study. Amplicon sizes are indicated at the top of the gel. Marker M is High Range DNA Ladder from Ampliqon (A610141)
COVERAGE OF AQ90
Illustration of the coverage of AQ90 High Fidelity DNA Polymerase. 10x AQ90 Buffer supports robust amplification of DNA targets with a GC content ranging from ~ 30 – 70 %. The addition of 2M Betaine Enhancer solution supports amplification of DNA targets with extremely high GC content. The coverage of AQ90 High Fidelity DNA Polymerase is illustrated against the coverage of AccuPol DNA polymerase from Ampliqon and Taq DNA Polymerase when using the 10x Ammonium Buffer.