HOT FIREPol® EvaGreen® HRM Mix (ROX)

High-Resolution Melt (HRM) analysis is a fast and cost-effective method for post-PCR analysis to detect single nucleotide polymorphisms (SNP genotyping), genetic mutations, and DNA sequence variations.

5x HOT FIREPol^® EvaGreen® HRM Mix is an optimized ready-to-use solution for HRM analysis, incorporating EvaGreen^® dye.

It comprises all the components necessary to perform qPCR and HRM Analysis: HOT FIREPol^® DNA Polymerase, ultrapure dNTPs, MgCl_2, and EvaGreen^® dye. The user simply needs to add water, template, and primers.

HOT FIREPol^® DNA Polymerase is activated by a 12 min incubation step at 95°C. This prevents the extension of non-specifically annealed primers and primer-dimers.

Applications

• High-Resolution Melt (HRM)
• SNP detection
• dye-based qPCR

Properties

Concentration: 5x

Hot-start: yes, initial activation in 12 min

Detection type: dye-based, includes EvaGreen^® intercalating dye

Reference dye: none

Compatible real-time instruments: Cyclers that do not require ROX reference dye. Check Your cycler!

Mix Components

HOT FIREPol^® DNA polymerase: chemically modified FIREPol^® DNA Polymerase enabling hot-start

5x EvaGreen^® qPCR buffer with 12. 5 mM MgCl₂, 1x PCR solution – 2.5 mM MgCl₂

dNTPs: dATP, dCTP, dGTP and dTTP

EvaGreen^® dye

Bovine serum albumin (BSA) to enhance qPCR reaction

EvaGreen Dye

EvaGreen^® is a DNA-binding dye with many features that make it a superior alternative to SYBR^® Green I for qPCR. Apart from having similar spectra, EvaGreen^® has three important features that set it apart from SYBR^® Green I: EvaGreen^® has much less PCR inhibition, is an extremely stable dye and has been shown to be non-mutagenic and non-cytotoxic. EvaGreen^® is compatible with all common real-time PCR cyclers – simply select the standard settings for SYBR® Green or FAM!