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以小麦根尖分生组织细胞核及染色体制备高分子量DNA 被引量：3

High Molecular Weight DNA Preparation from Nuclei or Chromosomes of Root Meristem of Wheat
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摘要 为了克服以叶片为材提取HMW DNA的局限性,优化了一套简便、实用的提取方法。该法以细胞周期同步化处理的根尖分生组织为材料,利用机械匀浆释放细胞核或中期染色体制备悬浮液,再以蔗糖密度梯度离心和流式分选技术分别分离细胞核和染色体,经去蛋白、酶切、透析,获得HMW DNA。经检测,来自200条根尖(10个胶块)的HMW DNA的浓度约为4～20ng/μL,而连接、转化后的BAC克隆平均插入片段超过100kb。证明该法适用于提取HMW DNA以构建BAC文库。 A simple and practical method was optimized to overcome the limitations of current method by which HMW DNA was prepared from leaf powder.Root tips of synchronized meristem were homogenized to make preparing suspension containing intact metaphase chromosomes and nuclei,which were further isolated by sucrose gradient centrifugation and flow sorting,respectively.Finally,HMW DNA was prepared with deproteinization,digestion and dialysis of nuclei and chromosomes.The results showed that concentration of the HMW D...
作者 郭东伟 佘茂云 李连城 陈耀峰 陈明 徐兆师 马有志
机构地区 西北农林科技大学农学院 中国农业科学院作物科学研究所/农业部遗传育种重点开放实验室
出处 《作物学报》 CAS CSCD 北大核心 2006年第12期1920-1923,共4页 Acta Agronomica Sinica
基金 西北农林科技大学青年教师基金
关键词 高分子量DNA 蔗糖密度梯度离心 细胞周期同步化 染色体流式分选 BAC High molecular weight DNA Sucrose gradient centrifugation Synchronization of cell cycle Chromosome flow sorting BAC
分类号 S512.1 [农业科学—作物学]
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参考文献12

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同被引文献42

  • 1寇姝燕,李文政,谭学林.烟草高分子量核DNA提取方法比较[J].中国农学通报,2006,22(3):64-66. 被引量：1
  • 2王贤智,张晓娟,周蓉,沙爱华,吴学军,蔡淑平,邱德珍,周新安.大豆重组自交系群体荚粒性状的QTL分析[J].作物学报,2007,33(3):441-448. 被引量：31
  • 3张洋,王志强,刘斌,张晓军,姜枫,相建海.DH10B菌株高效电转化条件探究[J].生物工程学报,2007,23(2):347-351. 被引量：10
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作物学报
2006年 第12期

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