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HRM Fast

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KAPA HRM FAST

Accurate detection of DNA sequence variations.

High Resolution Melt (HRM) analysis is a technique for fast, high-throughput post-PCR analysis of genetic mutations or variance in nucleic acid sequences. The KAPA HRM FAST Master Mix contains a novel DNA polymerase and buffer system, and EvaGreen® saturating dye optimized for maximum discrimination between sequence variants.

Product Highlights

   The Type I SNP rs12913832 is associated with expression of the OCA2 gene, and part of the eye color haplotype. Reactions also included 2.5 mM MgCl2, 0.2 µM of each primer and 10 ng of human genomic DNA. PCR and HRM was performed using a fast, 2-step cycling protocol (40 cycles) with 5 seconds denaturation (95°C) and 20 seconds annealing/extension (60°C). Competitor kits were used according to manufacturers’ instructions, with the same template DNA samples.

Maximize differences in sequence variants

  • Engineered to maximize differences in melting behavior
  • Improved sensitivity
    The Type IV SNP rs641805 is located on an intron of the dihydropyrimidine dehydrogenase (DPD) gene. Reactions also included 2.5 mM MgCl2, 0.2 μM of each primer and 10 ng of human genomic DNA. PCR and HRM was performed using a fast, 2-step cycling protocol (40 cycles) with 5 seconds denaturation (95°C) and 20 seconds annealing/extension (60°C).

Accurately detect Type IV SNPs

  • 95% detection levels, even when competitors fail
  • Reduced reaction times
           DNA extracts were prepared from buccal swabs obtained from 36 individuals using KAPA Express Extract DNA Extraction Kits (according to recommended protocols). DNA extracts were used directly in reactions also containing 2.5 mM MgCl2 and 0.2 μM of each primer. PCR and HRM was performed using a fast, 2-step cycling protocol (45 cycles) with 5 second denaturation (95°C) and 30 seconds annealing/extension (60°C).

 Go from cell to SNP quickly

  • Enables extraction, amplification and HRM in less than 2 hours when used in combination with the KAPA Express Extract Kit
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