ArticleThe Journal of molecular diagnostics : JMD2007
Unlabeled oligonucleotides as internal temperature controls for genotyping by amplicon melting.
Article in The Journal of molecular diagnostics : JMD, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed.
- Development and Application of a High-Resolution Melting Analysis with Unlabeled Probes for the Screening of Short-Tailed SheepAnimals : an open access journal from MDPI · 2022Article
- High Resolution Melting (HRM) for High-Throughput Genotyping-Limitations and Caveats in Practical Case Studies.International journal of molecular sciences · 2017Article
- SNP Marker Discovery in Pima Cotton (Genomics insights · 2016Article
- Article
- High-Resolution Melting Analysis as a Developed Method for Genotyping the PD Susceptibility Loci in LRRK2 Gene.Journal of clinical laboratory analysis · 2015Article
- Automated Classification and Cluster Visualization of Genotypes Derived from High Resolution Melt Curves.PloS one · 2015Article
- High-resolution melt as a screening method in autosomal dominant polycystic kidney disease (ADPKD).Journal of clinical laboratory analysis · 2014Article
- Article
- High-resolution melt analysis to detect sequence variations in highly homologous gene regions: application to CYP2B6.Pharmacogenomics · 2013Article
- High-resolution melting analysis for SNP genotyping and mapping in tetraploid alfalfa (Medicago sativa L.).Molecular breeding : new strategies in plant improvement · 2012Article
- High-resolution melting molecular signatures for rapid identification of human papillomavirus genotypes.PloS one · 2012Article
- Detection of the JAK2 mutation in myeloproliferative neoplasms by asymmetric PCR with unlabeled probe and high-resolution melt analysis.Journal of clinical laboratory analysis · 2011Article
- Development of high-resolution melting (HRM) analysis for population studies of Fascioloides magna (Trematoda: Fasciolidae), the giant liver fluke of ruminants.Parasitology research · 2011Article
- Automated DNA extraction, quantification, dilution, and PCR preparation for genotyping by high-resolution melting.Journal of biomolecular techniques : JBT · 2010Article
- Nucleotide extension genotyping by high-resolution melting.The Journal of molecular diagnostics : JMD · 2010Article
- High-resolution melting analysis, a simple and effective method for reliable mutation scanning and frequency studies in the ACADVL gene.Journal of inherited metabolic disease · 2010Article
- High-throughput identification and quantification of Candida species using high resolution derivative melt analysis of panfungal amplicons.The Journal of molecular diagnostics : JMD · 2010Article
- Multiplex amplicon genotyping by high-resolution melting.Journal of biomolecular techniques : JBT · 2009Article
- Description and validation of high-throughput simultaneous genotyping and mutation scanning by high-resolution melting curve analysis.Human mutation · 2009Article
- High resolution melting applications for clinical laboratory medicine.Experimental and molecular pathology · 2008Review
Corrections and comments
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Authors and funding
9 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Amplicon melting is a closed-tube method for genotyping that does not require probes, real-time analysis, or allele-specific polymerase chain reaction. However, correct differentiation of homozygous mutant and wild-type samples by melting temperature (Tm) requires high-resolution melting and closely controlled reaction conditions. When three different DNA extraction methods were used to isolate DNA from whole blood, amplicon Tm differences of 0.03 to 0.39 degrees C attributable to the extractions were observed. To correct for solution chemistry differences between samples, complementary unlabeled oligonucleotides were included as internal temperature controls to shift and scale the temperature axis of derivative melting plots. This adjustment was applied to a duplex amplicon melting assay for the methylenetetrahydrofolate reductase variants 1298A>C and 677C>T. High- and low-temperature controls bracketing the amplicon melting region decreased the Tm SD within homozygous genotypes by 47 to 82%. The amplicon melting assay was 100% concordant to an adjacent hybridization probe (HybProbe) melting assay when temperature controls were included, whereas a 3% error rate was observed without temperature correction. In conclusion, internal temperature controls increase the accuracy of genotyping by high-resolution amplicon melting and should also improve results on lower resolution instruments.
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