Enhanced primer selection and synthetic amplicon templates optimize high-resolution melting analysis of single-nucleotide polymorphisms in a large population

Carol M. Bruzzone, Patrick S. Tawadros, Lisa A. Boardman, Clifford J. Steer

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

Aims: High-resolution melting (HRM) screening and scanning for single-nucleotide polymorphisms (SNPs) afford the advantages of a quicker, less expensive, and less demanding option compared to other methods for sequence analysis. The evaluation of large populations of patients for multiple SNPs in a high-throughput manner is the next phase in individualized medicine. Results: We demonstrated that Tm profiles can be generated from gDNA samples that clearly differentiate homozygous ancestral, homozygous SNP, and heterozygous genotypes, while identifying samples of unique outcome without the cumbersome processes of normalization, temperature shifting, and difference plot generation. Conclusions: Through expanded primer selection criterion and inclusion of a cloning fragment length double-stranded DNA sequence-specific control template, we are now able to generate additional data via HRM melt domains that are greatly simplified, while considering both the peak melt temperature and profile.

Original languageEnglish (US)
Pages (from-to)675-680
Number of pages6
JournalGenetic Testing and Molecular Biomarkers
Volume17
Issue number9
DOIs
StatePublished - 2013

ASJC Scopus subject areas

  • Genetics(clinical)

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