Assessing high-resolution melt curve analysis for accurate detection of gene variants in complex DNA fragments.
Mutation detection has until recently relied heavily on the use of gel-based methods that can be both time consuming and difficult to design. Non gel-based systems are therefore important to increase simplicity and improve turn-around-time without compromising assay sensitivity and accuracy, especially in the diagnostic/clinical setting. In this study, we assessed the latest of the non gel-based methods, namely high resolution melt (HRM) curve analysis. HRM is a closed-tube method that incorporates a saturating dye during DNA amplification followed by a monitoring of the change in fluorescence as the DNA duplex is denatured by an increasing temperature. We assessed ten amplicons derived from eight genes, namely SERPINA1, CXCR7, MBL, VDR, NKX3A, NPY, TP53 and HRAS using two platforms, the LightScanner
|Authors||Tindall, E.; Petersen, D.C.; Woodbridge, P.; Schipany, K.; Hayes, V. M.:|
|Publisher Name||HUMAN MUTATION|
|URL link to publisher's version||http://www3.interscience.wiley.com/journal/122255765/abstract?CRETRY=1&SRETRY=0|
|OpenAccess link to author's accepted manuscript version||https://publications.gimr.garvan.org.au/open-access/10031|