AQ97 Hot Start High Fidelity DNA Polymerase 2x Master Mix
AQ97 Hot Start High Fidelity DNA Polymerase 2x Master Mix (AQ97 HS HiFi 2x Master Mix) is convenient alternative to AQ97 Hot Start High Fidelity DNA Polymerase (AQ97 HS HiFi), displaying the same set of features. The master mix is based on AQ97 Hot Start High Fidelity DNA Polymerase, which is a thermostable, chimeric DNA polymerase created specifically for low-bias, high fidelity amplification of a vast range of amplicons.
The antibody-mediated inactivation of the polymerase and exonuclease functions increases specificity of the master mix, as well as enabling room temperature setup.
AQ97 HS HiFi 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 setup
Reaction set-up at room temperature
Low error rate
Low-bias amplification
Amplification of long amplicons (up to 11 kb)
High elongation rate
DESCRIPTION
AQ97 HS HiFi 2x Master Mix is a ready to use 2x reaction mix composed of AQ97 Hot Start High Fidelity DNA Polymerase and an optimized buffer system including dNTPs and magnesium chloride, allowing robust amplification on DNA target with low to high GC content and long DNA targets.
AQ97 Hot Start High Fidelity DNA Polymerase exhibits both 5'→3' DNA polymerase activity and 3'→5' proofreading exonuclease activity, enabling accurate correction of base-pair mismatches. The attached DNA-binding domain enhances processivity, while the antibody-mediated inactivation ensures specificity even at room temperature setup. Together, these features yield an enzyme that consistently delivers low-error, specific, and fast results with long-range capacity and low-bias amplification.
For targets longer than 11 kb we recommend the stand-alone AQ97 HS HiFi. The ability to adjust reaction conditions, such as dNTPs and enzyme concentration, enables it to amplify DNA targets up to 18 kb.
Addition of 1-2 M Betaine Enhancer Solutions is recommended when amplifying difficult amplicons, such as GC-rich targets, long targets, or those with complex secondary structures.
FIDELITY
Polymerase fidelity refers to the accuracy with which a DNA polymerase synthesizes a complementary strand during replication. The fidelity rate is influenced by factors such as the specific polymerase used, the buffer system, and the quality of the template DNA.
The fidelity of AQ97 HiFi was examined with a novel NGS-based method, counting the number of base substitutions within a PCR product. Data is available here.
LOW-BIAS AMPLIFICATION
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. Marker M: Iqon PCR Ladder from Ampliqon (A610641).
LONG-RANGE AMPLIFICATION
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 from Ampliqon (A610141).
Product info
AQ97 Hot Start High Fidelity DNA Polymerase 2x Master Mix
