Abstract
To the Editor: Genotyping by amplicon melting is a minimalistic approach to genotyping, requiring only PCR, a fluorescent DNA dye, and single-color acquisition of a melting curve in a closed tube. Multiplex genotyping for the F5 1 [coagulation factor V (proaccelerin, labile factor)] Leiden (1691G>A), F2 [coagulation factor II (thrombin)] 20210G>A, and MTHFR [methylenetetrahydrofolate reductase (NAD(P)H)] 1298A>C and 677C>T thrombophilia mutations distinguishes up to 81 (34) combinations in solution without physical processing (1). Furthermore, we previously suggested that it might be possible to detect additional rare variants within the sequences of the quadruplex amplicons by a combination of melting temperature ( T m) and shape discrimination enabled by high-resolution melting analysis. Amplified heterozygotes have a complex melting curve shape consisting of 2 homoduplexes (wild type and mutant) and 2 heteroduplexes that are influenced by the class of single-nucleotide polymorphism and other variants …
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
