KAPA Plant PCR Kit 500U HALO MPR-96 Microplate Reader

KAPA2G™ Fast Multiplex PCR Mix 500rxns

Product Description

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.

KAPA2G Fast Multiplex PCR Kits are designed for Fast Multiplex PCR and offer faster and more efficient Multiplex PCR than competitor enzymes based on wild-type Taq DNA polymerase. The increased speed and processivity of the engineered KAPA2G Fast HotStart DNA Polymerase results in a significant decrease in cycling times, whilst still maintaining the ability to efficiently amplify difficult fragments. The uniquely formulated KAPA2G Fast Multiplex Mix facilitates primer annealing and highly specific amplification of a wide range of amplicon sizes and GC contents, resulting in more even amplification of all target fragments.

The KAPA2G Fast Multiplex PCR Kit is ideally suited for end-point, Fast Multiplex PCR of multiple DNA fragments, ranging in size from 50 – 1'500bp. Up to 30 different primer pairs may be combined into a single assay using the protocols provided in the Technical Sheet. The kit is ideally suited for:

typing of transgenic organisms
amplification of microsatellites
typing and detection of pathogens
amplification of multiple DNA fragments for SNP genoptyping

Results obtained with KAPA2G Fast HotStart “Multiplex PCR Mix” in multiplex PCR of twenty human amplicons. Reactions (25μl) contained 1X Multiplex PCR Mix, 0.2μM of each primer and a 5-fold dilution series of human genomic DNA (250ng – 0.4ng per reaction, as indicated above each lane). KAPA2G Fast HotStart reactions were performed with 3 minutes initial denaturation at 95°C, followed by 30 cycles of denaturation at 95°C (15 seconds), annealing at 60°C (30 seconds) and extension at 72°C (30 or 60 seconds). Total cycling time for KAPA2G Fast HotStart reactions was 1h22min (30 seconds extension) or 1h37min (60 seconds extension). Reactions were analysed by agarose gel electrophoresis (2% agarose-TBE gel), and visualised by ethidium bromide staining.