KAPA HRM FAST PCR Kits

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Engineered for fast and 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 PCR Kit contains a convenient, ready-to-use master mix designed for the high performance detection of DNA sequence variations.

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. Kits are ideally suited for:


NEW! Application Guide: Introduction to High Resolution Melt Analysis. Download here>>

Code Description Kit Contents Qty Unit Prices
KK4201 KAPA HRM FAST PCR Kit (1 mL) For 100 x 20 µL reactions. Supplied with separate 1 x 1.6 mL 25 mM MgCl2. login for pricing
KK4202 KAPA HRM FAST PCR Kit (5 mL) For 500 x 20 µL reactions. Supplied with separate 2 x 1.6 mL 25 mM MgCl2. login for pricing
KK4203 KAPA HRM FAST PCR Kit (2 x 5 mL) For 1000 x 20 µL reactions. Supplied with separate 4 x 1.6 mL 25 mM MgCl2. login for pricing

 

 

NEW! Application Guide: Introduction to High Resolution Melt Analysis. Download here>>


Product Description

The KAPA HRM FAST PCR Kit is a convenient, ready-to-use master mix designed for the high performance detection of DNA sequence variations by High Resolution Melt (HRM) analysis. The kit contains a novel DNA polymerase, engineered via a process of molecular evolution, for fast and efficient DNA amplification in the presence of high concentrations of intercalating fluorescent dyes. KAPA HRM FAST PCR Kits also contain EvaGreen®, a next-generation, saturating fluorescent dye which selectively binds to double-stranded DNA. In contrast to SYBR® Green I dye, EvaGreen® can be used at higher concentrations without PCR inhibition and shows equal binding affinity for GC-rich and AT-rich regions. The combination of an engineered DNA polymerase and EvaGreen® dye enables amplification and discrimination of even the most challenging sequence differences (i.e. Type IV) without sequence preference.


Product Applications

KAPA HRM FAST PCR Kits are optimized for endpoint HRM analysis and are not recommended for quantitative PCR. Kits are ideally suited for:

  • SNP genotyping
  • Mutation discovery
  • Species identification and genotyping
  • DNA fingerprinting
  • Genetic association studies


Accurate SNP genotyping with maximum sensitivity and speed.

KAPA HRM FAST PCR Kits are engineered to maximize differences in melting behavior between sequence variants. Kits contain a novel DNA polymerase and buffer system that allows for fast PCR cycling and improved sensitivity. Single nucleotide polymorphism (SNP) genotpying is the most common application for HRM. In this example, KAPA HRM FAST is compared to other HRM kits when genotyping Type I SNP rs12913832 (G/A).

The Type I SNP rs12913832 is associated with expression of the OCA2 gene, and part of the eye color haplotype. Reactions (20 µl) contained 1X KAPA HRM FAST PCR Master Mix, 2.5 mM MgCl2, 0.2 µM of each primer (targeting a 60 bp amplicon surrounding the SNP of interest) and 10 ng of human genomic DNA (all three genotypes, confirmed by sequencing). PCR and HRM was performed with the Corbett Rotor-Gene™ 6000 instrument, using a fast, 2-step cycling protocol (40 cycles) with 5 sec denaturation (95 °C) and 20 sec annealing/extension (60 °C). Competitor kits were used according to manufacturers’ instructions, with the same template DNA samples.

 

Genotyping of the Type I SNP rs12913832 (G/A) with the KAPA HRM FAST PCR Kit.

Genotyping of the Type I SNP rs12913832 (G/A) with the QIAGEN Type-it HRM PCR Kit.

Genotyping of the Type I SNP rs12913832 (G/A) with the Bio-Rad SsoFast™ EvaGreen® SuperMix.

Fast and reproducible detection of Type IV SNPs.

The Type IV SNP rs641805 is located on an intron of the dihydropyrimidine dehydrogenase (DPD) gene, which catalyzes the reduction of uracil and thymine and is associated with the degradation of certain chemotherapeutic drugs. Reactions (20 μl) contained 1X KAPA HRM FAST PCR Master Mix, 2.5 mM MgCl2, 0.2 μM of each primer (targeting a 124 bp amplicon surrounding the SNP of interest) and 10 ng of human genomic DNA (all three genotypes, confirmed by sequencing). PCR and HRM was performed with the Corbett Rotor-GeneTM 6000 instrument, using a fast, 2-step cycling protocol (40 cycles) with 5 sec denaturation (95 °C) and 20 sec annealing/extension (60 °C). The ABI MeltDoctor HRM Master Mix was used according to the manufacturer’s instructions, with the same template DNA samples. ABI MeltDoctorTM is unable to differentiate the Type IV SNP, while KAPA HRM FAST detects the SNP in 95% of the samples assayed.

 

Genotyping of the Type IV SNP rs641805 (A/T) with the KAPA HRM FAST PCR Kit.

Genotyping of the Type IV SNP rs641805 (A/T) with the ABI MeltDoctorTM HRM Master Mix.

From cell to SNP: Rapid genotyping directly from buccal swabs.

The combination of the KAPA Express Extract DNA Extraction Kit (containing a novel thermostable protease) and the KAPA HRM FAST PCR Kit enables extraction, amplification and HRM in less than 2 hours. DNA extracts were prepared from buccal swabs obtained from 36 individuals using KAPA Express Extract DNA Extraction Kits (according to recommended protocols). DNA extracts (1 μl of each) were used directly in 20 μl KAPA HRM FAST reactions containing 1X KAPA HRM FAST Master Mix, 2.5 mM MgCl2 and 0.2 μM of each primer (targeting a 75 bp amplicon surrounding the SNP of interest). PCR and HRM was performed with the Corbett Rotor-GeneTM 6000 instrument, using a fast, 2-step cycling protocol (45 cycles) with 5 sec denaturation (95 °C) and 30 sec annealing/extension (60 °C).

 

Genotyping of the Type II SNP rs12896399 (G/T), part of the eye color haplotype, with KAPA Express Extract and KAPA HRM FAST.

Licensing Information

Certain applications of this product are covered by patents issued to parties other than Kapa Biosystems and applicable in certain countries. Purchase of this product does not include a license to perform any such applications. Users of this product may therefore be required to obtain a patent license depending upon the particular application and country in which the product is used.

Use of this product is covered by one or more of the following US patents and corresponding patent claims outside the US: 5,994,056, 6,171,785, and 5,928,907 (claim numbers 12-24, 27-28). The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser’s own internal research. No right under any other patent claim (such as apparatus or system claims in US Patent No. 6,814,934) and no right to perform commercial services of any kind, including without limitation reporting the results of purchaser’s activities for a fee or other commercial consideration, is conveyed expressly, by implication, or by estoppel. This product is for research use only. Diagnostic uses under Roche patents require a separate license from Roche. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.

Kapa Biosystems Inc. is licensed by Biotium Inc. to sell reagents containing EvaGreen dye for use in real-time PCR, for research purposes only.

Licensed under U.S. Patent nos. 5,338,671 and 5,587,287 and corresponding patents in other countries.

The purchase of this product includes a limited, non-transferable license under specific claims of U.S. Patent Nos. 6,174,670, 6,569,627 and 5,871,908, owned by the University of Utah Research Foundation or Evotec Biosystems GmbH and licensed to Idaho Technology, Inc. and Roche Diagnostics GmbH, to use only the enclosed amount of product according to the specified protocols. No right is conveyed, expressly, by implication, or by estoppel, to use any instrument or system under any claim of U.S. Patent Nos. 6,174,670, 6,569,627 and 5,871,908, other than for the amount of product contained herein.

EvaGreenTM is a registered trademark of Biotium, Inc. Rotor-GeneTM is a trademark of Corbett Research.

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DocumentTypeDownload
Introduction to High Resolution Melt Analysis Guide Guide Download
KAPA HRM FAST MSDS Material Safety Data Sheet Download
KAPA HRM FAST Brochure Product Brochure Download
KAPA HRM FAST TDS Technical Data Sheet Download