三肽囊素(Bursin)同功单链抗体(ScFv)的构建研究 Ⅰ.鸡脾脏组织细胞总RNA提取方法研究  

Extraction of Total RNA of Tissue Cells of Chicken Spleen

作　　者：邬向东[1] 曲悦[1] 谌南辉[1] 

机构地区：[1]江西农业大学动科院,江西南昌330045

出　　处：《江西农业大学学报》2003年第2期289-291,共3页Acta Agriculturae Universitatis Jiangxiensis

基　　金：江西省自然科学基金资助项目(0 13 0 0 2 3 );江西省科技厅农业科技攻关项目

摘　　要：分离纯净、完整的细胞总RNA对于分子克隆的实验十分重要 ,但自然界中RNA酶广泛存在且稳定性很高 ,RNA极易被RNA酶污染而降解。快速提取RNA可以减少RNA酶污染的机会而成为RNA提取的首选方法。选用 3种快速RNA提取法 (方法I、II、III) ,发现方法III最好 ,电泳可见 3个RNA条带清晰无降解 ,纯度也最高 ;方法II较差 ,虽然操作时间最短 ,但却易降解 ,而且电泳时多出一条明显的DNA条带 ;方法I也能提取到较好的RNA ,但纯度不如方法III。Isolation of intact and pure cellular total RNA is very important to molecular cloning. However RNases are rich and stable in the nature, so RNA is liable to degradation caused by contamination from exogenous RNases. Rapid extraction of RNA reduces the chances of contamination from RNases. Three methods for direct and rapid extraction of total RNA are introduced in this paper, namely, method Ⅰ, methodⅡ and method Ⅲ. Method Ⅲ is the best. The RNA extracted with method Ⅲ is not degraded and of the highest degree of purity; three bands (28s, 18s and 5s) are clearly seen when checked with ethidiam-agrose gel electrophoresis. Method Ⅱ is not as good, a DNA band is observed in electrophoresis; and the RNA is degraded. Method Ⅰ can also extract RNA with comparatively high quality, but its purity is lower than that of the RNA extracted with method Ⅲ.

关 键 词：三肽囊素同功单链抗体 RNA 提取方法 组织细胞 RNA酶 

分 类 号：Q813.2[生物学—生物工程] Q522