TY - JOUR
T1 - High-resolution melt analysis for SNP discovery, linkage mapping and analysis of DNA methylation
AU - Croxford, Adam E.
AU - Lopez, C. M. R.
AU - Wilkinson, Michael J.
N1 - Lopez, C. M. R., Croxford, A. E., Wilkinson, M. J. (2008). High-resolution melt analysis for SNP discovery, linkage mapping and analysis of DNA methylation Comparative Biochemistry and Physiology A-Molecular & Integrative Physiology, Abstracts, Annual Meeting of the Society-for-Experimental-Biology, Marseille, FRANCE, JUL 06-10, 2008, 150, (3), S49-S50
PY - 2008/7/1
Y1 - 2008/7/1
N2 - High Resolution Melt analysis (HRM) is a closed-tube method of genotyping that does not require use of fluorescent probes, fragment fractionation or amplicon sequence information. Recent advancements in florescent-detection instruments (such as the Corbett Rotor-Gene 6000) and the use of fully saturating intercalating dyes have made HRM analysis considerably more sensitive. The flexibility of the system allows it to be adapted for a wide range of uses including SNP genotyping, mutation detection, screening for loss of heterozygosity, DNA fingerprinting, characterization of haplotype blocks, species classification, somatically acquired mutations studies, linkage and physical mapping, and DNA methylation analysis. Here, we describe the first use of high-resolution melt analysis to generate STS markers based on Single Nucleotide Polymorphisms (SNPs) and microsatellite length polymorphisms for use in linkage mapping, using white lupin (Lupinus albus, x = 25) as a case study. The described strategy is rapid and low-cost, and circumvents the need for labeled primers or amplicon fractionation. We also illustrate the use of HRM analysis for the detection and/or quantification of the presence of, and relative abundance of, methylated nucleic acid bases within the double-stranded molecule without any prior chemical modification of the target DNA.
AB - High Resolution Melt analysis (HRM) is a closed-tube method of genotyping that does not require use of fluorescent probes, fragment fractionation or amplicon sequence information. Recent advancements in florescent-detection instruments (such as the Corbett Rotor-Gene 6000) and the use of fully saturating intercalating dyes have made HRM analysis considerably more sensitive. The flexibility of the system allows it to be adapted for a wide range of uses including SNP genotyping, mutation detection, screening for loss of heterozygosity, DNA fingerprinting, characterization of haplotype blocks, species classification, somatically acquired mutations studies, linkage and physical mapping, and DNA methylation analysis. Here, we describe the first use of high-resolution melt analysis to generate STS markers based on Single Nucleotide Polymorphisms (SNPs) and microsatellite length polymorphisms for use in linkage mapping, using white lupin (Lupinus albus, x = 25) as a case study. The described strategy is rapid and low-cost, and circumvents the need for labeled primers or amplicon fractionation. We also illustrate the use of HRM analysis for the detection and/or quantification of the presence of, and relative abundance of, methylated nucleic acid bases within the double-stranded molecule without any prior chemical modification of the target DNA.
U2 - 10.1016/j.cbpa.2008.04.035
DO - 10.1016/j.cbpa.2008.04.035
M3 - Article
SN - 1095-6433
VL - 150
SP - S49-S50
JO - Comparative Biochemistry and Physiology - Part A: Molecular and Integrative Physiology
JF - Comparative Biochemistry and Physiology - Part A: Molecular and Integrative Physiology
IS - 3
T2 - Abstracts, Annual Meeting of the Society-for-Experimental-Biology, JUL 06-10
Y2 - 1 January 2008 through 1 January 2008
ER -