BACKGROUND High-resolution DNA melting analysis with saturation dyes for either mutation scanning of PCR products or genotyping with unlabeled probes has been reported. However, simultaneous PCR product scanning and probe genotyping in the same reaction has not been described. METHODS Asymmetric PCR was performed in the presence of unlabeled oligonucleotide probes and a saturating fluorescent...

Hereby we present evaluation of high-resolution melting for mutation scanning applied to the cystic fibrosis transmembrane conductance regulator gene. High resolution melting was used for mutation scanning of selected samples derived from cystic fibrosis patients with a known cystic fibrosis transmembrane conductance regulator genotype. We tested 19 different disease-causing cystic fibrosis tra...

The melting transition of Xe monolayers adsorbed on a single-crystal exfoliated graphite substrate has been studied by high-resolution synchrotron x-ray scattering. At temperatures slightly above the melting transition the fluid phase has a high degree of orientational order. The results are discussed in the context of current theories of two-dimensional melting including the effects of the sub...

High-resolution melting curve analysis is a closed-tube fluorescent technique that can be used for genotyping and heteroduplex detection after polymerase chain reaction. We applied this technique at the HLA-A locus and suggest that this method can be used as a rapid, inexpensive screen between siblings prior to living-related transplantation. At any locus, there are seven general cases of share...

BACKGROUND Multiplexed amplicon melting is a closed-tube method for genotyping that does not require probes, real-time analysis, asymmetric PCR, or allele-specific PCR; however, correct differentiation of homozygous mutant and wild-type samples by melting temperature (T(m)) analysis requires high-resolution melting analysis and controlled reaction conditions. METHODS We designed 4 amplicons b...

High-resolution melting of DNA is a simple solution for genotyping, mutation scanning and sequence matching. The melting profile of a PCR product depends on its GC content, length, sequence and heterozygosity and is best monitored with saturating dyes that fluoresce in the presence of double-stranded DNA. Genotyping of most variants is possible by the melting temperature of the PCR products, wh...

High resolution melting (HRM) is a novel, closed-tube, post-PCR technique allowing genomic researchers to easily analyze genetic variations in PCR amplicons. This technical note describes general steps of setting up HRM-based PCR assays, with a special focus on ways how to optimize procedures for gene scanning experiments. The novel High Resolution Melting (HRM) technique enables researchers to...