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For Research Use Only. Not for use in diagnostic procedures.
Overview
KAPA2G Fast PCR Kits contain a second-generation (2G) DNA polymerase enzyme engineered for higher processivity and speed, offering significantly faster extension rates than wild-type Taq polymerase. In addition to speed, KAPA2G Fast PCR Kit provides higher yields and sensitivity than competitor enzymes across a broad range of targets.
Resources
Product highlights
High speed and performance compared to competitor kits
Reduced cycling times, up to 75%*
Extension times as low as 1 sec per kilobase
Broad coverage of both AT- and GC-rich targets
View human genomic DNA amplification data showing reduced cycling time and better PCR performance using KAPA2G Fast PCR Kit compared to competitor kits, and data showing reduced reaction times using HotStart KAPA2F Fast DNA polymerase compared to HotStart wild type Taq enzyme.
Figure 1. Amplification of 5 human gene fragments using KAPA2G Fast HotStart or competitor hot-start Taq formulations. Reactions (25 µL) contained 5 ng human genomic DNA and 0.5 units (KAPA2G Fast HotStart Polymerase enzyme and Competitor I Kit) or 0.625 units (Competitor A and Q) enzyme. For amplicons with a GC content >65% (lanes 2 and 3), 7.5% DMSO was included in reactions. A 3-step cycling profile (35 cycles) with 15 sec denaturation (95˚C) and 15 sec annealing (60˚C) per cycles was used for all enzymes. The extension (72˚C) was 1 sec/cycle for KAPA2G Fast HotStart enzyme and 60 sec/cycle for competitor enzymes. The total reaction time for each enzyme is indicated.
Figure 2. Total PCR times for the generation of different sized amplicons for human genomic DNA (≤3.5 kb) and lambda (≤5 kb) using wild-type Taq DNA polymerase and KAPA2G Fast DNA Polymerase. The amount of time that can be saved using protocols based on wild-type Taq is limited by the extension rate of the enzyme. KAPA2G Fast DNA Polymerase is based on a second-generation polymerase with an ability to synthesize DNA faster than wild-type Taq or other DNA polymerases. Reaction times are based on a 35-cycle program using the cycling profile recommended by each kit manufacturer.
View performance data generated using HotStart KAPA2F Fast PCR Kit showing amplification of a 1.3 kb fragment of the macrophage stimulating factor 1 receptor gene.
Figure 3. Amplification of a 1.3 kb fragment of the macrophage stimulating 1 receptor gene from a 10-fold dilution series of human genomic DNA using KAPA2G Fast HotStart. 0.5 units of enzymes were used per 25 µL reaction. A standard 3-step cycling profile (35 cycles) with a total extension time of 15 sec/kb per cycle was used.
< 100 ng for genomic DNA, cDNA
< 10 ng for less complex DNA
Kits can be stored for up to 18 months at -20˚C.
Standard kits include KAPA2G Fast (5 U/μL), KAPA2G Buffer A (5X, with 1.5 mM MgCl2 at 1X) or KAPA2G Buffer M (5X, MgCl2-free), and MgCl2(25 mM). Kits are available with and without dNTP Mix (10 mM each).
ReadyMix formulations (2X) of KAPA2G Fast and KAPA2G Fast HotStart DNA Polymerase are also available. These contain DNA polymerase, reaction buffer (with and without dye), and MgCl2 (at a final concentration of 1.5 mM).
