碱性强弱染色非变性聚丙烯酰胺凝胶对微卫星DNA检测效果的影响  

Detecting of Microsatellite DNA by Native PAGE Stained in Strong or Weak Alkali Solution

作　　者：汪虹英[1] 

机构地区：[1]西南科技大学材料科学与工程学院,四川绵阳621010

出　　处：《西南科技大学学报》2011年第3期84-87,共4页Journal of Southwest University of Science and Technology

基　　金：四川省教育厅重点项目(08ZA010)资助

摘　　要：非变性聚丙烯酰胺凝胶电泳常用于分离不同分子质量和空间构象的DNA或RNA等生物大分子,由于分辨率可以达到2个碱基对,因此非变性聚丙烯酰胺凝胶对于微卫星DNA有很好的检测效果。利用非变性聚丙烯酰胺凝胶电泳对黄牛微卫星DNA的检测表明:碱性强弱的控制对于染色的效果和效率有显著的影响。弱碱性环境下,具有背景浅、条带明显和检测灵敏度高的优势,但是染色过程较复杂繁琐,适合于少量样品的精细检测;在强碱性环境下,对于微卫星DNA的灵敏度相对弱碱性环境下降低,但是染色过程简便、快捷且成本相对较低,适合于大规模样品的检测。Native polyacrylamide gel electrophoresis （PAGE） is used to isolate such biological macromol- eeules as DNA or RNA based on the differences of their molecular weight and spatial configuration. As the nucleic acids divergent from two base pairs could be isolated by this kind of eleetrophoresis, PAGE should be one of effective methods to isolate microsatellite DNA. In this work, microsatellite DNA of cattle was detected by using native PAGE and the staining of gel with strong or weak alkali had significant effect on the results and effectiveness of detection. In the weak alkali solution, the background of the gel is bright, the DNA lanes are distinct and the detection is sensitive. Whereas, the process of this weak alkali stained PAGE is complex and time-consuming. Therefore, this method will apply to the sensitive detection of DNA samples with small quantities. In the strong alkali solution, the sensitiveness of PAGE is comparatively reduced, while the process of this strong alkali stained PAGE is much more simplified, cheap and convenient. Therefore, this method will apply to the detection of DNA samples with large quantities.

关 键 词：碱性环境 非变性聚丙烯酰胺凝胶 微卫星DNA 

分 类 号：Q78[生物学—分子生物学]