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Assessing high-resolution melt curve analysis for accurate detection of gene variants in complex DNA fragments.

Abstract

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

Type Journal
ISBN 1098-1004 (Electronic)
Authors Tindall, E.; Petersen, D.C.; Woodbridge, P.; Schipany, K.; Hayes, V. M.:
Publisher Name HUMAN MUTATION
Published Date 2009-06-01
Published Volume 30
Published Issue 6
Published Pages 876-883
Status Published in-print
DOI 10.1002/humu.20919
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