gb HRM PCR Master Mix

gb HRM PCR Master Mix contains hot-start Taq DNA polymerase, reaction buffer, dNTP, MgCl2, fluorescent dye (with detection in FAM/Sybr channel) and other components needed for PCR amplification with subsequent HRM analysis. For mixing of reaction mixture only add primers and template.

Description of the product

gb HRM PCR Master Mix is intended for amplification of DNA with subsequent High Resolution Melting (HRM) analysis. HRM analysis is an efficient melt curve analysis method for genotyping, mutation scanning prior to sequencing analysis, DNA mapping, species identification or DNA methylation analysis etc. It is based on dissociation behavior of nucleic acids due to increasing temperature gradient. With the use of a double stranded DNA fluorescent dye it allows to detect even a single base change in the sequence of an amplified PCR product. No additonal instrumentation is required; HRM analysis can be performed on any real-time PCR cycler simply by adding a Melt Curve step analysis with high resolution directly after a PCR amplification. This product is intended for research use only.

Cat. No. Product No. of reactions Price
3017 gb HRM PCR Master Mix 100
Request

1 tube contains reagents to provide 100 PCR reactions (20 μl volume of each reaction).

Parameters of the product

  • Master Mix is 2x concentrated.
  • One vial contains 1 ml of Master Mix (this amount is sufficient for 100 reactions of 20 µl). PCR Master Mix contains hot-start Taq DNA polymerase, reaction buffer, dNTP, MgCl2, fluorescent dye (with detection in FAM/Sybr channel) and other components needed for PCR amplification with subsequent HRM analysis.
  • PCR Master Mix does not contain any passive reference dye.
  • If the real-time PCR cycler requires normalization with a passive reference dye, it can be added to the reaction mixture separately (see Passive Reference Dye, Cat. No. 3010 – on request)

 

Amplification protocol

Initial denaturation 95 °C 3 min
Denaturation 95 °C 5 s 30-50 cycles
Annealing+elongation (+fluorescence acquiring) 55-65 °C 10 s 30-50 cycles
Extension 72 °C 20 s 30-50 cycles
Denaturation 95 °C 2 min
HRM analysis/Melt Curve (+fluorescence acquiring) 50 °C 20 s
HRM analysis/Melt Curve (+fluorescence acquiring) 70-95 °C 5s rise by 0,1-0,2 °C

 

gb PCR Master Mix by application
gb HRM
end-point PCR, common PCR amplification
real-time PCR without probes

real-time PCR with hydrolysis probes
real-time PCR with LNA probes
real-time PCR with hybridization probes
real-time PCR with High Resolution Melting Analysis

real-time PCR with low DNA samples
real-time PCR with inhibited sampels

Useful links
Recommended products
Recommended products
gb Reverse Transcription Kit

gb Reverse Transcription Kit gb Reverse Transcription Kit

Reverse Transcription Kit is intended for reverse transcription using enzyme MMLV reverse transcriptase. This enzyme converts RNA to cDNA (complementary DNA) in one step (without denaturation), you only have to add your template RNA.

More