Multiplex PCR is a challenging application that typically requires more optimization than standard, single amplicon PCR assays. The key to successful Multiplex PCR is the ability to define a single set of reaction parameters (reagent concentrations and cycling parameters) that allows for all primers to anneal with high specificity to their target sequences and be extended with the same efficiency. Primer design, as well as the enzyme and buffer system, are critical factors in this challenge.
The KAPA2G Fast Multiplex PCR Kit offers faster and more efficient Multiplex PCR than competitor enzymes based on wild-type Taq. KAPA2G Fast Multiplex Kit contains the KAPA2G Fast HotStart DNA Polymerase, an engineered DNA Polymerase designed specifically for high performance Fast PCR. In the HotStart formulation, the enzyme is combined with a proprietary antibody that inactivates the enzyme until the initial denaturation step. This eliminates spurious amplification products resulting from non-specific priming events during reaction setup and initiation, and increases overall reaction efficiency. Together with a uniquely formulated buffer, the KAPA2G Fast Multiplex PCR Kit is the product of choice for Multiplex PCR.
Key performance advantages are the following:
- Total reaction times 30 - 70% shorter than protocols based on wild-type Taq or hot start formulations thereof.
- A uniquely formulated multiplex buffer that improves annealing specificity, resulting in more even amplification of target fragments.
- Improved sensitivity, as a result of higher reaction efficiency. This means less template is required.
- A straightforward process for conversion or optimization of existing Multiplex PCR assays.
KAPA PROBE FAST qPCR Kits are recommended for Multiplex qPCR using sequence specific TaqMan® probes, FRET probes, and molecular beacons.