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https://hdl.handle.net/2440/9959
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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 1095-9130 |
Statement of Responsibility: | 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 Cancer Tumor Silencing Bisulfite Single-strand conformation analysis Single-strand conformation polymorphism Genomic sequencing p16 |
Rights: | © 2002 Elsevier Science (USA). All rights reserved. |
DOI: | 10.1016/S1046-2023(02)00065-8 |
Description (link): | http://www.elsevier.com/wps/find/journaldescription.cws_home/622914/description#description |
Published version: | http://dx.doi.org/10.1016/s1046-2023(02)00065-8 |
Appears in Collections: | Aurora harvest Medicine publications |
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