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Creating localized DNA double-strand breaks with microirradiation.


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タイトル: Creating localized DNA double-strand breaks with microirradiation.
著者: Suzuki, Keiji / Yamauchi, Motohiro / Oka, Yasuyoshi / Suzuki, Masatoshi / Yamashita, Shunichi
発行日: 2011年 2月
出版者: Nature Publishing Group
引用: Nature Protocols, 6(2), pp.134-139; 2011
抄録: We describe a protocol for creating localized DNA double-strand breaks (DSBs) with minimal requirements that can be applied in cell biology and molecular biology. This protocol is based on the combination of 5-bromo-2'-deoxyuridine (BrdU) labeling and ultraviolet C (UVC) irradiation through porous membranes. Cells are labeled with 10 μM BrdU for 48-72 h, washed with Ca(2+)- and Mg(2+)-free PBS(-), covered by polycarbonate membranes with micropores and exposed to UVC light. With this protocol, localized DSBs are created within subnuclear areas, irrespective of the cell cycle phase. Recruitment of proteins involved in DNA repair, DNA damage response, chromatin remodeling and histone modifications can be visualized without any specialized equipment. The quality is the same as that obtained by laser microirradiation or by any other focal irradiation. DSBs become visible within 30 min of UVC irradiation.
記述: without figures
キーワード: DNA double strand breaks / micro-irradiation / DNA damage-induced foci / local damage / DNA damage response / DNA repair
URI: http://hdl.handle.net/10069/25685
ISSN: 17542189
DOI: 10.1038/nprot.2010.183
PubMed ID: 21293454
権利: © 2011 Nature America, Inc. All rights reserved.
資料タイプ: Journal Article
原稿種類: author
出現コレクション:110 学術雑誌論文

引用URI : http://hdl.handle.net/10069/25685

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