斑点杂交_____]斑点杂交_；糖基转移酶；基因

基于尼龙膜的点杂交条件的建立及优化

http://www.100md.com 《实用医技杂志》 2006年第4期

斑点杂交,,]斑点杂交,；糖基转移酶；基因,1材料和方法,2结果,参考文献：

     [关键词]斑点杂交；糖基转移酶；基因

    [摘要] 目的：进行基于尼龙膜的点杂交条件的建立及优化。方法：采用带正电荷的尼龙膜对斑点杂交的每一步骤进行优化，包括预杂交液，杂交液，洗涤和封闭液等及各步反应时间，并采用最终确立的条件检测糖基转移酶基因多肽：N乙酰氨基半乳糖转移酶2(ppGalNAcT2)在K562、NB4、NIH3T3和SGC7901细胞株中的差异表达以验证膜杂交条件的可行性。结果：建立并优化了膜斑点杂交的条件，本底清晰，结果可靠。结论：建立的斑点杂交技术经济简便，切实可行。

    Establishment and Optimization of Dot Blot Technique

    SHAO Xuejun1,WU Shiliang2

    (1.Soochow University Affiliated Children's Hospital，Suzhou Jiangsu 215003, China；2.Department of Biochemistry & Molecular Biology,Medical School,Soochow University，Suzhou, Jiangsu 215007, China)

    Abstract:Objective To establish and optimize the dot blot technique.Methods We used the positively charged nylon membranes to optimize experiments for the dot blot including prehybridization solution，hybridization solution，washing buffer，blocking solutionetal，and the times of every step.In order to validate its feasibility，we analyzed the expression of ppGalNAcT2 in different tumor cell lines by dot blot assay using the established hybridizationsystem.Results we established and optimized the dot blot technique which had reliable results and low background.Conclusion The established dot blot technique is practicable and economical. ......

百拇医药网 http://www.100md.com/html/DirDu/2006/05/21/97/67/45.htm

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