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Type: Journal article
Title: Screening for and analysis of methylation differences using methylation-sensitive single-strand conformation analysis
Author: Dobrovic, A.
Bianco-Miotto, T.
Tan, L.
Sanders, T.
Hussey, D.
Citation: Methods, 2002; 27(2):134-138
Publisher: Academic Press Inc
Issue Date: 2002
ISSN: 1046-2023
Statement of
Alexander Dobrovic, Tina Bianco, Lor Wai Tan, Tanya Sanders and Damian Hussey
Abstract: Methylation-sensitive single-strand conformation analysis (MS-SSCA) is a method of screening for methylation changes at CpG sites in a region of DNA. After bisulfite modification, the region of interest is amplified using primers specific for bisulfite-modified sequences. The amplified products are denatured and run on a nondenaturing polyacrylamide gel. The sequence differences caused by methylation lead to the formation of different secondary structures (conformers) with different mobilities. MS-SSCA is a convenient and rapid method for screening large numbers of samples for methylation. Individual bands can readily be isolated and sequenced allowing more detailed analysis of methylation changes. In this article, we present a protocol for MS-SSCA and outline strategies for the design of primers for amplifying bisulfite-modified DNA sequences.
Keywords: Methylation
Single-strand conformation analysis
Single-strand conformation polymorphism
Genomic sequencing
Rights: © 2002 Elsevier Science (USA). All rights reserved.
DOI: 10.1016/S1046-2023(02)00065-8
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