SensiMix HRM Kit


Features and Benefits Cost effective: method is less expensive and as sensitive as probe-based methods, making it ideal for large scale genotyping projects Simple and reproducible: powerful genotyping can be performed by non-geneticists in any laboratory with access to an HRM capable real-time PCR machine Sensitive: detects class 4 (A/T) SNP mutations Convenient: with optimized protocols, reliable assays can quickly and reliably be established, even with genomic loci that are difficult to amplify Instrument Compatibility Roche LightCycler® 480 and the Corbett Rotor-GeneTM 6000 (See product selection table). Description High Resolution Meltcurve (HRM) analysis characterizes nucleic acid samples based on their dissociation behavior. It combines the principle of intercalating dyes, meltcurve analyses and the application of specific statistical analyses HRM uses the fundamental property of the separation of the two strands of DNA with heat (melting), and the monitoring of this melting with a fluorescent dye. The SensiMix HRM Kit employs a 3rd generation saturating dye that does not inhibit PCR at high concentrations and has been validated for use on the Roche LightCycler® 480 and the Corbett Rotor-Gene™ 6000. Melting temperature of a dsDNA target depends on GC content, length, and sequence. Due to the high sensitivity of HRM dyes, even a single base change will induce differences in the melting curve, and consequently in fluorescence. Main applications of HRM include Single Nucleotide Polymorphisms (SNPs) genotyping, epigenetics (DNA methylation analysis), zygosity testing (DNA mapping and DNA fingerprinting) and gene scanning (search for the presence of unknown variations in PCR amplicons). Volume QT805-02: 250 x 25µl Reactions: 5 x 625µl QT805-05: 500 x 25µl Reactions: 10 x 625µl QT805-20: 2000 x 25µl Reactions: 40 x 625µl Components 2x Reaction mix consisting of: Heat-activated DNA polymerase Ultra-pure dNTPs MgCl₂ (6mM) Proprietary buffers and stabilisers Plus separate vials of: EvaGreen™ dye 50mM MgCl₂ Kit Size The kit size is based on a 25µl final reaction volume. Storage Conditions 6 months at -20°C. Avoid exposure of the EvaGreen™ to light. Product Citations 1. Chana W-F. et al. Journal of Microbiological Methods 77(3): 326-329 (2009) 2. Andriantsoanirina V. et al. Journal of Microbiological Methods 78(2): 165-170 (2009) 3. FitzGerald L.M. et al. The Prostate 68(13): 1373-1379 (2008) 4. Helen E. et al. Clinical Chemistry 53(11): 1960-1975 (2007)