qPCR Green Master HighROX with UNG 2X | Detecção por EvaGreen para Genotipagem / High Resolution Melt (HRM) - Cellco
2X Master mix for real-time qPCR Green-fluorescent DNA Stain

For in vitro use only!

Envio:
Shipped on blue ice

Condições de armazenamento:
Store at -20 °C
(Avoid freeze/thaw cycles, store in dark). Store at 4 °C for up to 3 months possible.

Concentração:
2 x conc.

Validade:
12 months

Forma:
Liquid

Spectroscopic Properties:
λ_exc 500 nm (bound to DNA); λ_em 530 nm (bound to DNA)

Kit contents:
qPCR Green Master (red cap)
qPCR Pol, dATP, dCTP, dGTP, dUTP, EvaGreen®, 1 µM ROX, reaction buffer with KCl, (NH₄)₂SO₄, MgCl₂ and stabilizers

ROX reference dye:
The qPCR GreenMaster highROX with UNG contains 500 nM ROX passive reference dye in the final assay. The dye does not take part in the PCR reaction but allows to normalize for non-PCR related signal variation and provides a baseline in multiplex reactions.

EvaGreen® Fluorescent DNA Stain:
EvaGreen® Fluorescent DNA Stain is a superior 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. The high quantum yield, excellent stability and lowest inhibition toward PCR makes it the ideal fluorophore in real-time PCR applications and a superior replacement for the widely used SYBR® Green I dye. To perform the EvaGreen-based assay simply select the optical setting for SYBR® Green or FAM on the detection instrument.

Description:
qPCR GreenMaster is designed for the quantitative real-time analysis of DNA samples using the fluorescent DNA stain EvaGreen®. The fluorescent dye in the master mix intercalates into the amplification product during the PCR process and enables the rapid analysis of target DNA without the need to synthesize sequence-specific labeled probes. It provides an easy-to-handle and powerful tool for quantification of sample DNA in a broad dynamic range of up to 6 orders of magnitude with exceptional sensitivity and precision. The Master contains all reagents required for qPCR (except template and primer) in a premixed 2x concentrated ready-to-use solution. The high specificity and sensitivity of the mix is achieved by an optimized hot-start polymerase. Its activity is blocked at ambient temperature and switched on automatically at the onset of the initial denaturation. The thermal activation prevents the extension of nonspecifically annealed primers and primer-dimer formations at low temperatures during PCR setup. The mix contains UNG (Uracil-N-Glycosylase) and dUTP instead of dTTP to eliminate carry-over contamination of DNA from previous PCR reactions. The UNG treatment at the onset of thermal cycling removes uracil residues from dU-containing DNA and prevents it from serving as template. The reaction chemistry of the kit is optimized for instruments that are compatible with the evaluation of a high ROX reference signal.