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PubMed 19768593


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Title: Zn(II)-cyclen polyacrylamide gel electrophoresis for SNP detection.

Authors: Emiko Kinoshita-Kikuta, Eiji Kinoshita, Tohru Koike

Journal, date & volume: Methods Mol. Biol., 2009 , 578, 169-82

PubMed link: http://www.ncbi.nlm.nih.gov/pubmed/19768593


Abstract
We introduce a method for the detection of single-nucleotide polymorphisms (SNPs) by polyacrylamide gel electrophoresis (PAGE) with an additive Zn2+-cyclen complex (cyclen is 1,4,7,10-tetraazacyclododecane), called "Zn2+-cyclen-PAGE." The method is based on the difference in mobility of mutant DNA (of the same length) in PAGE, which is due to Zn2+-cyclen binding to thymine bases accompanying a total charge decrease and a local conformation change of target DNA. In combination with a polymerase chain reaction based heteroduplexing technique, the method is more accurate than many other conventional PAGE-based methods, as shown by clear gel-shifting bands because of the separation of heteroduplex and homoduplex DNAs. We demonstrate SNP mapping and heterozygosity screening in a human cardiac sodium channel gene (i.e., SCN5A) that relates to inherited arrhythmia syndromes using Zn2+-cyclen-PAGE.