ArticleJournal of biomolecular techniques : JBT2009
Multiplex amplicon genotyping by high-resolution melting.
Article in Journal of biomolecular techniques : JBT, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed.
- Proof of concept, randomized, placebo-controlled study of the effect of simvastatin on the course of age-related macular degeneration.PloS one · 2013Trial
- Demonstration of Neural Networks to Reconstruct Temperatures from Simulated Fluorescent Data Toward Use in Bio-microfluidics.International journal of thermophysics · 2022Article
- Use of Machine Learning with Temporal Photoluminescence Signals from CdTe Quantum Dots for Temperature Measurement in Microfluidic Devices.ACS applied nano materials · 2020Article
- SLAM-MS: Mutation scanning of stem-loop amplicons with TaqMan probes by quantitative DNA melting analysis.Scientific reports · 2020Article
- Extreme PCR Meets High-Speed Melting: A Step Closer to Molecular Diagnostics "While You Wait".Clinical chemistry · 2019Article
- A Novel Multiplex HRM Assay to Detect Clopidogrel Resistance.Scientific reports · 2017Article
- HRM and SNaPshot as alternative forensic SNP genotyping methods.Forensic science, medicine, and pathology · 2017Article
- High-Resolution Melting Analysis as a Developed Method for Genotyping the PD Susceptibility Loci in LRRK2 Gene.Journal of clinical laboratory analysis · 2015Article
- Rapid and inexpensive body fluid identification by RNA profiling-based multiplex High Resolution Melt (HRM) analysis.F1000Research · 2013Article
- Rapid multiplex high resolution melting method to analyze inflammatory related SNPs in preterm birth.BMC research notes · 2012Article
- High-resolution melting analysis as a powerful tool to discriminate and genotype Pseudomonas savastanoi pathovars and strains.PloS one · 2012Article
- 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 curve analysis of DNA sequence alterations of various sizes.Pathology oncology research : POR · 2010Article
- Rapid genetic analysis of x-linked chronic granulomatous disease by high-resolution melting.The Journal of molecular diagnostics : JMD · 2010Article
- High resolution melting analysis for gene scanning.Methods (San Diego, Calif.) · 2010Article
Corrections and comments
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Authors and funding
4 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
High-resolution amplicon melting is a simple method for genotyping that uses only generic PCR primers and a saturating DNA dye. Multiplex amplicon genotyping has previously been reported in a single color, but two instruments were required: a carousel-based rapid cycler and a high-resolution melting instrument for capillaries. Manual transfer of capillaries between instruments and sequential melting of each capillary at 0.1 degrees C/s seriously limited the throughput. In this report, a single instrument that combines rapid-cycle real-time PCR with high-resolution melting [LightScanner-32 (LS-32), Idaho Technology, Salt Lake City, UT] was used for multiplex amplicon genotyping. The four most common mutations associated with thrombophilia, F5 (factor V Leiden 1691G>A), F2 (prothrombin 20210G>A), and methylenetetrahydrofolate reductase (MTHFR; 1298A>C and 677C>T) were genotyped in a single homogeneous assay with internal controls to adjust for minor chemistry and instrument variation. Forty temperature cycles required 9.2 min, and each capillary required 2.2 min by melting at 0.3 degrees C/s, 3x the prior rate. Sample volume was reduced from 20 microl to 10 microl. In a blinded study of 109 samples (436 genotypes), complete concordance with standard assays was obtained. In addition, the rare variant MTHFR 1317T>C was genotyped correctly when present. The LS-32 simplifies more complex high-resolution melting assays by reducing hands-on manipulation, total time of analysis, and reagent cost while maintaining the resolution necessary for multiplex amplicon genotyping.
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19568454PMC2700465What OpenQuestion holds
Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.