Detection of the Gua/Cyt-to-Cyt/Gua mutation in a Gua/Cyt-lined sequence using Zn2+–cyclen polyacrylamide gel electrophoresis
Analytical Biochemistry 380 巻 1 号
122-127 頁
2008-09-01 発行
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種類 :
全文
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タイトル ( eng ) |
Detection of the Gua/Cyt-to-Cyt/Gua mutation in a Gua/Cyt-lined sequence using Zn2+–cyclen polyacrylamide gel electrophoresis
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作成者 |
Kinoshita-Kikuta Emiko
Yoshimoto Miki
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収録物名 |
Analytical Biochemistry
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巻 | 380 |
号 | 1 |
開始ページ | 122 |
終了ページ | 127 |
抄録 |
We have previously reported a method for the detection of single-nucleotide polymorphisms by polyacrylamide gel electrophoresis (PAGE) with an additive Zn2+–cyclen complex (cyclen = 1,4,7,10-tetraazacyclododecan), called Zn2+–cyclen–PAGE. The method is based on the difference in mobility of mutant DNA (in 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 heteroduplexing technique, the method is more accurate, as shown by clear gel-shifting bands. However, the question remains of whether the Gua/Cyt-to-Cyt/Gua mutation, which is far apart from the Thy/Ade base (i.e., in a Gua/Cyt-lined sequence), can be detected by this Thy-dependent method. In this study, we determined the potency of Zn2+–cyclen–PAGE for the detection of the Gua/Cyt-to-Cyt/Gua single substitutions in some artificial Gua/Cyt-lined sequences derived from a human cardiac sodium channel gene, SCN5A. All Gua/Cyt-to-Cyt/Gua substitutions in the 28-set samples tested, which are 1 to 10 bases away from the nearest Thy/Ade, were successfully detected by designing DNA fragments of the appropriate length.
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著者キーワード |
Guanine
Cytosine
SNP
Mutation
Heteroduplex
Zn2+–cyclen
Electrophoresis
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NDC分類 |
医学 [ 490 ]
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言語 |
英語
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資源タイプ | 学術雑誌論文 |
出版者 |
Academic Press Inc.
Elsevier Science
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発行日 | 2008-09-01 |
権利情報 |
Copyright (c) 2008 Elsevier Inc.
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出版タイプ | Author’s Original(十分な品質であるとして、著者から正式な査読に提出される版) |
アクセス権 | オープンアクセス |
収録物識別子 |
[ISSN] 0003-2697
[DOI] 10.1016/j.ab.2008.05.040
[NCID] AA00524867
[DOI] http://dx.doi.org/10.1016/j.ab.2008.05.040
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