©2009 UTMB Molecular Genomics Core (MGC)

ABOUT High resolution melt (hrm)
Service Information
High Resolution Melt (HRM) Technology Description
HRM FAQs
 

The Molecular Genomics Core (MGC) now offers High-Resolution Melt (HRM) as a service to our clients.  HRM is a novel  post-PCR method, enabling genomic researchers to analyze genetic variations (SNPs, mutations, methylations) in PCR amplicons.  The most important high-resolution melting application is gene scanning – the search for the presence of unknown variation in PCR amplicons prior to or as an alternative to sequencing. 
Mutations in PCR products are detectable by HRM because they influence the shape of DNA melting curves. A combination of new-generation DNA dyes, high-end instrumentation and sophisticated analysis software allows detection of these changes and to derive information about the underlying sequence constellation.

Samples should be submitted in the form of genomic DNA either frozen or as an ethanol precipitate.  Once received, sample quantity will be determined via the NanoDrop®.  Once samples pass QC, HRM analysis will be carried out on our Roche LightCycler 480.  A customized report will be generated.

HRM Advantages
HRM provides high specificity and sensitivity.  Furthermore, it allows processing of high sample numbers more conveniently and at much lower cost than traditional, non-homogeneous (gel-based) mutation screening methods (e.g., dHLPC) that require amplicons to be screened for variants on a separate instrument after PCR. 
With HRM melting, amplicons can be screened for unknown sequence variants with a single high-resolution dye.  Allele-specific primers or probes to target specific variants are not needed, and variants can be detected regardless of their position within the fragment.

Service Features & Options

  • Sample Quantitation via NanoDrop® ND-1000
  • Assay design  
  • Assay Optimization (magnesium /primer concentration)
  • Easy to understand data report

Service Information
Technology Description – HRM
FAQs –HRM

Useful links
RT-QPCR vs Traditional PCR

Roche LightCycler 480

To schedule an appointment or to discuss experimental design, send an email, or call 409-772-6349.


© 2009 University of Texas
301 University Blvd.; Galveston, TX 77555