Detecção por EvaGreen para Genotipagem/ High Resolution Melt (HRM)

JBS qPCR GreenMaster Series is designed for quantitative real-time analysis of DNA samples including High Resolution Melting (HRM) curve analysis. The mixes contain all reagents required for qPCR (except template and primers) in a premixed 2x concentrated ready-to-use solution. The mixes are recommended for routine PCR applications, high throughput PCR or genotyping and provide an improved specificity and sensitivity when amplifying low-copy-number targets or working with complex backgrounds.
The fluorescent DNA stain EvaGreen® intercalates into the amplification product during the PCR process and allows the direct quantification of target DNA without the need of sequence-specific labeled probes (i.g. TaqMan® Probes).

High Resolution Melting (HRM) analysis is a powerful tool for detection of mutations, polymorphisms and epigenetic differences in DNA samples. It is a fast and cost effective alternative to other genotyping technologies.

UNG (Uracil-N-Glycosylase) eliminates carry-over contamination of DNA from previous PCR reactions. An UNG treatment at the onset of thermal cycling removes uracil residues from dU-containing DNA and prevents it from serving as template.

ROX reference dye does not take part in the PCR reaction but allows to normalize for non-PCR related signal variation.

EvaGreen® Fluorescent DNA Stain is a DNA intercalator dye specially developed for DNA analysis applications including real-time PCR (qPCR) and high-resolution DNA melting curve analysis (HRM). Upon binding to DNA, the non-fluorescent dye becomes highly fluorescent while showing no detectable inhibition to the PCR process. The dye is extremely stable both thermally and hydrolytically, providing convenience during routine handling.

SYBR® Green é uma marca registrada pela Invitrogen Corporation, Carlsbad, California, USA.