The KAPA2G Fast PCR Kit is designed for the conversion of existing PCR assays into Fast PCR assays for the purpose of reducing the total reaction time by up to 70%, without compromising reaction performance or having to invest in specialized PCR consumables or instrumentation.

The superior speed and performance of the KAPA2G Fast PCR Kit is attributable to KAPA2G Fast DNA Polymerase, a novel, engineered enzyme derived from Taq DNA polymerase using Kapa Biosystems’ molecular evolution platform.

Designed specifically for speed and performance, the KAPA2G Fast PCR kit offers Fast PCR that is not merely based on the reduction of cycling times, but on the intrinsic ability of the KAPA2G Fast DNA polymerase to synthesize DNA at a much faster rate.

Product Applications

KAPA2G Fast PCR Kits are designed for the fast amplification of DNA fragments up to 5 kb in standard end-point PCR assays, using reaction volumes up to 25 ul. Amplicons generated with these kits are suitable for routine downstream applications, including restriction enzyme digestion, cloning and sequencing.

Product Performance

Extreme Speed and High Performance

KAPA2G Fast DNA Polymerase is capable of dramatically reduced extension times without compromising performance. Extension rates as low as 10 sec/kb for amplicons 2-5 kb in size, 7.5 kb/sec for 1-2 kb amplicons and 1 sec/kb for amplicons <1 kb may be used while retaining exceptional yield and dynamic range.

Using the same amount of enzyme per reaction and a standard protocol with 1 min/kb extension per cycle, Taq polymerase is unable to match the yield and sensitivity of KAPA2G Fast DNA Polymerase for the amplification of large fragments. As extension times are reduced, the performance of Taq polymerase deteriorates significantly, thereby limiting the time that can be saved using fast PCR protocols based on wild-type Taq. For the amplification of a 5 kb fragment, the significantly faster extension rate of the KAPA2G Fast enzyme translates into a saving of 50-67% without any further optimization of reaction parameters.

For fragments of 1 kb or less, a total extension time of 1 second is sufficient for reproducible amplification from as little as a single target copy using the KAPA2G Fast PCR kit.

Amplification of a 1 kb fragment from a 10-fold dilution series of human genomic DNA using the KAPA2G Fast PCR kit, using 1 sec total extension time per cycle. All reactions contained 0.5 units of enzyme per 25 µl reaction. Cycling was performed on the G-Storm GS1 thermocycler with a fast block, using the following cycling profile: 95°C (2 min), 35 cycles of 95°C (15 sec), 64oC (15 sec), 72°C (1 sec) and a final extension of 30 sec (72°C).

By reducing extension time only, the total reaction time was reduced by 49%. With further optimization and fast thermocyclers, short amplicons may be routinely amplified in about 20 min using the KAPA2G Fast PCR Kit.

KAPA2G Fast PCR Kit Components

KAPA2G Fast DNA Polymerase (5 U/µl)
5x KAPA2G Buffer A with MgCl₂
5x KAPA2G Buffer B with MgCl₂
25 mM MgCl₂
KAPA dNTP Mix (10 mM each nucleotide)

For more information https://www.kapabiosystems.com/product/kapa2g/