HRMFASTKB

KAPA HRM FAST

• Synonym(s): PCR
• UNSPSC Code: 41106300
• NACRES: NA.55

Properties

• shelf life: ≤12 mo.
• packaging: kit of 1 mL (100 x 20 μL rxn; KK4201); kit of 10 mL (1000 x 20 μL rxn; KK4203); kit of 5 mL (500 x 20 μL rxn; KK4202)
• manufacturer/tradename: Roche
• technique(s): PCR: suitable
• storage temp.: −20°C

Description

General description

High Resolution Melt (HRM) analysis is a technique for fast, high-throughput post-polymerase chain reaction (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. The KAPA HRM FAST PCR Kit is a convenient and ready to use master mix.

Application

KAPA HRM FAST has been used for:

• Single nucleotide polymorphism (SNP) genotyping
• High resolution melt (HRM) assay -Mutation discovery (gene scanning)
• Heterozygosity screening
• DNA fingerprinting
• DNA methylation analysis
• Amplicon or allele amplification
• Genome editing screening for single colonies

Features and Benefits

Maximize differences in sequence variants

• Engineered to maximize differences in melting behavior.
• Improved sensitivity.

Accurately detect Type IV SNPs

• 95% detection levels, even when competitors fail.
• Reduced reaction times.

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.

Quick Notes:

• KAPA HRM FAST PCR Kits contain EvaGreen, a saturating double-stranded DNA dye optimized for HRM analysis.
• The 2X Master Mix contains a buffer system optimized for HRM analysis and a novel DNA polymerase to enhance amplification efficiency. The Master Mix does not contain MgCl₂.
• Template DNA from different samples should be prepared using the same method and eluted or diluted in the same buffer (ideally 10 mM Tris-HCl, pH 8.5).
• The optimal amount of template DNA is between 20 ng and 100 pg per 20 μL reaction.
• To maximize the sensitivity of the assay, use the lowest concentration of primers that can be used without compromising the efficiency of the PCR reaction (0.1 μM – 0.3 μM of each primer).
• The concentration of MgCl₂ can greatly affect the annealing of heterozygotes and melting of the amplicons during HRM. The recommended starting concentration is 2.5 mM MgCl₂, which can be adjusted as required. 1.5 – 3.5 mM final MgCl₂ is recommended for most HRM assays.
• Instrument gain settings may have to be adjusted for maximum sensitivity using KAPA HRM FAST PCR Kits.

Quality

All kit components are subjected to stringent functional quality control, are free of detectable contaminating exo and endonuclease activity, and meet strict requirements with respect to DNA contamination.

Preparation Note

Always ensure that components have been fully thawed and thoroughly mixed before use. KAPA HRM FAST qPCR Master Mix (2X) may not freeze solidly, even when stored at -20°C.
The EvaGreen dye contained in KAPA HRM FAST qPCR Master Mix (2X) is light sensitive. Exposure to direct light for an extended period of time will result in loss of fluorescent signal intensity.
KAPA HRM FAST qPCR Master Mix (2X) is stable through 30 freeze-thaw cycles. Ensure that all reagents are stored protected from light at -20°C when not in use. When protected from light, reagents are stable in the dark at 4°C for at least one week and may be stored at this temperature for short-term use, provided that they do not become contaminated with microbes and/or nucleases.

Other Notes

For Research Use Only. Not for use in diagnostic procedures.

Legal 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.

EvaGreen is a registered trademark of Biotium, Inc.
Rotor-Gene is a trademark of Corbett Research.

EvaGreen is a registered trademark of Kapa Biosystems

Kit Components Only

• KAPA HRM FAST MasterMix (2X)
• KAPA MgCl₂ 25 mM

Safety Information

• Storage Class Code: 12 - Non Combustible Liquids
• WGK: WGK 1
• Flash Point(F): does not flash
• Flash Point(C): does not flash