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, MgCl2, 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!
