| 棉花gDNA体细胞染色体FISH技术 |
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| 引用本文： | 王春英,王坤波,王文奎,李懋学,宋国立,崔荣霞,黎绍惠,张香娣,张家明.棉花gDNA体细胞染色体FISH技术[J].棉花学报,1999(2). |
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| 作者姓名： | 王春英 王坤波 王文奎 李懋学 宋国立 崔荣霞 黎绍惠 张香娣 张家明 |
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| 作者单位： | 中国农业科学院棉花研究所，北京林业大学园林学院，北京大学生命科学院 |
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| 摘 要： | 介绍了棉花基因组ＤＮＡ（ｇｅｎｏｍｅＤＮＡ，简为ｇＤＮＡ）体细胞染色体荧光原位杂交〔ＦＩＳＨ〕的技术流程，并着重分析和讨论了影响试验结果的关键因素，包括染色体和探针的变性条件、染色体的蛋白酶Ｋ处理技巧等。试验中作为靶ＤＮＡ的体细胞染色体采用棉属异源四倍体种海岛棉；探针和封阻均采用ｇＤＮＡ，材料是棉属二倍体种Ａ染色体组（Ａｇｅｎｏｍｅ）的棉种（亚洲棉和草棉）和Ｄ染色体组（Ｄｇｅｎｏｍｅ）的棉种（瑟伯氏棉、雷蒙德氏棉、戴维逊氏棉等），分别交互使用。试验结果比较理想，获得良好的ＦＩＳＨ片子，而且重复性好。
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| 关 键 词： | 棉花 基因组DNA 体细胞染色体 荧光原位杂交 |
Protocol of Cotton FISH of Somatic Chromosomes with gDNA as Probes |
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| WANG Chun ying ,WANG Kun bo ,WANG Wen kui ,LI Mao xue ,SONG Guo li ,CUI Rong xia ,LI Shao hui ,ZhANG Xiang di ,ZHANG Jia ming Cotton Research Institute,CAAS, Key Laboratory of Cotton Genetical Improvement,Mini.Protocol of Cotton FISH of Somatic Chromosomes with gDNA as Probes[J].Cotton Science,1999(2). |
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| Authors: | WANG Chun ying WANG Kun bo WANG Wen kui LI Mao xue SONG Guo li CUI Rong xia LI Shao hui ZhANG Xiang di ZHANG Jia ming Cotton Research Institute CAAS Key Laboratory of Cotton Genetical Improvement Mini |
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| Institution: | WANG Chun ying 1,WANG Kun bo 1,WANG Wen kui 2,LI Mao xue 3,SONG Guo li 1,CUI Rong xia 1,LI Shao hui 1,ZhANG Xiang di 1,ZHANG Jia ming 4 1 Cotton Research Institute,CAAS, Key Laboratory of Cotton Genetical Improvement,Mini |
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| Abstract: | Accessions used in the study included Gossypium barbadense as target DNA(somatic chromosomes), gDNA (genome DNA) of species of A genome including G.arboreum and G.herbaceum, of D genome including G.davidsonii, G.thurberi and G.raimondii, as probe DNA and/or block DNA, respectively. This paper described protocol of the FISH (Fluorescence in situ hybridization), with discussions emphasized on the key procedures of denaturation of chromosomes and probes, of proteinase K was used in treatment to chromosome preparations. |
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| Keywords: | cotton FISH somatic chromosomes gNDA |
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