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[1]李晓玲,李克秀,赵洪锟,等.野生大豆核基因组Mb级DNA的制备与酶切[J].大豆科学,2011,30(01):33-36,40.[doi:10.11861/j.issn.1000-9841.2011.01.0033]
　LI Xiao-ling,LI Ke-xiu,ZHAO Hong-kun,et al.Preparing and Digesting Megabase-size Nuclear DNA of Glycine soja Genome[J].Soybean Science,2011,30(01):33-36,40.[doi:10.11861/j.issn.1000-9841.2011.01.0033]

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野生大豆核基因组Mb级DNA的制备与酶切

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第30卷期数: 2011年01期页码: 33-36,40 栏目: 出版日期: 2011-02-25

Title:: Preparing and Digesting Megabase-size Nuclear DNA of Glycine soja Genome

文章编号:: 1000-9841（2011）01-0033-04

作者:: 李晓玲¹; 2; 李克秀²; 赵洪锟¹; 董英山¹; 1．吉林省农业科学院生物技术研究中心，吉林长春130033;
2. 长春工业大学化学与生命科学学院, 吉林长春 130012

Author(s):: LI Xiao-ling¹; 2; LI Ke-xiu²; ZHAO Hong-kun¹; DONG Ying-shan¹; 1.Biotechnology Research Centre, Jinlin Academy of Agricultural Sciences， Changchun 130033；
2.School of Chemistry and Life Science Changchun University of Technology， Changchun 130012, Jilin, China

关键词:: 野生大豆; Mb级DNA; 脉冲电泳; 基因组可转化人工染色体(TAC) 文库

Keywords:: Glycine soja; Megabase-size DNA; Pulsed field gel electrophoresis; TAC Genomic library

分类号:: Q781

DOI:: 10.11861/j.issn.1000-9841.2011.01.0033

文献标志码:: A

摘要:: 以野生大豆黄化幼苗为材料提取其细胞核DNA，经LMP包埋并用蛋白酶K裂解其中的核蛋白后，采用脉冲电泳回收2 Mb左右细胞核DNA。用HindⅢ对回收细胞核DNA进行部分酶切并用脉冲电泳回收酶切后的DNA片段，经电洗脱、浓缩和透析后DNA溶液浓度可达10 ng·μL^-1。连接转化检测结果表明：该DNA可用于后续野生大豆基因组可转化人工染色体(TAC) 文库的构建和基因组分析。

Abstract:: Nuclear DNA was extracted from etiolated seedlings of Glycine soja. Generally, the nuclei were embeded in low-melting-point agarose pulgs, digested with proteinase K and depurated with pulsed field gel electrophoresis to yield about two-Megabase-size DNA. The concentration of DNA solution could be up to 10 ng·μL^-1 after partial digestion by HindⅢ, elution by pulsed field gel electrophoresis,concentration and dialysis. The result of examination by ligation and eletroporation showed that the DNA obtained by the method was suitable for consequent construction of genomic library of Glycine soja.

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备注/Memo

项目基金：农业部野生植物资源保护资助项目（2010）；农业部CWRC资助项目（CWRC－20080205）；国家重点基础研究发展计划“973计划”资助项目（2007CB116205）；国家转基因专项资助项目（2008ZX08004-004）。

?第一作者简介：李晓玲（1971-），女，博士，副教授，主要从事植物生物技术与植物分子生物学研究。E-mail：lxl5275278@163.com。

更新日期/Last Update: 2014-09-11