Zinc(II)-cyclen polyacrylamide gel electrophoresis for detection of mutations in short Ade/Thy-rich DNA fragments

• Kinoshita-Kikuta, Emiko

• Kinoshita, Eiji

• Harada, Naoko

• Koike, Tohru

抄録

We describe an improved gel-based method with an additive Zn2+-cyclen complex (cyclen, 1,4,7,10-tetraazacyclododecane), Zn2+-cyclen-PAGE, for mutation detection in DNA fragments by PCR that contain more than 65% Ade/Thy bases and fewer than 100 base pairs (bp). Existing techniques have a problem in analyzing such short Ade/Thy-rich fragments because the duplexes are disrupted and are not detectable due to binding of Zn2+-cyclen to Thy bases. In this strategy using a PCR primer with a Gua/Cyt-lined sequence attached at its 5'-end, we successfully detected a mutation in an 86-bp Ade/Thy-rich region of the BRCA1 gene from formalin-fixed paraffin-embedded breast cancer-tissue sections.

収録刊行物

• Analytical Biochemistry

  Analytical Biochemistry 408 (2), 348-350, 2011-01-15

  Academic Press Inc Elsevier Science

キーワード

• Zn2+-cyclen
• PAGE
• SNP
• Mutation
• FFPE
• BRCA1

詳細情報

• CRID: 1050859215939376896
• NII論文ID: 120002753732
• NII書誌ID: AA00524867
• Web Site: http://ir.lib.hiroshima-u.ac.jp/00030873
• 本文言語コード: en
• 資料種別: journal article
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