几种植物材料中总RNA的提取

Isolation of Total RNA from Different Plant Species

用改良前后的异硫氰酸胍一步法、Trizol法和CTAB-异硫氰酸胍法,从多糖和色素较少的豌豆、水稻、构树和华南蕨绿叶,色素较多的红绒球、蟛蜞菊和红背桂花叶,以及富含多糖的香蕉果肉等几种植物材料中提取RNA。发现一步法对豌豆、水稻和蟛蜞菊叶片总RNA的提取均有很好的效果。Trizol法也有好的效果,但如果提取缓冲液的pH达6.0,提取的RNA便有DNA的污染,且该污染随pH增加而增加。对于富含多糖的香蕉果肉,一步法和CTAB异硫氰酸胍法的效果均不理想;改良后的一步法中增加了乙醚去除多糖并获得了较好的效果。本文还提出了判断RNA的质量的标准。

By modified and un-modified single step guanidinium thiocyanate, Trizol and CTAB-guanidinium thiocyanate methods, total RNA were isolated from several species, such as from green leaves of pea, rice, Broussonetia Papyrifera, Cycloru Parasiticus, that contained little polysaccharide and pigment, from Calliandra Haematocephala, Wedelia chinensis(Osb.) Merr. and Excoecaria cochinensis Lour., that contained a lot of pigment, and from banana flesh, a material was rich in polysaccharide. Single step method worked well when total RNA in pea, rice, Broussonetia Papyrifera, Cycloru Parasiticus and Wedelia chinensis (Osb.) Merr. were isolated. Trizol method was a good method, but DNA always contaminated RNA preparation when pH of extraction solution was or greater than 6.0, and DNA/RNA ratio increased as pH increase. All single step method and CTAB method did not work well when banana total RNA was isolated, unless modified single step method was used, in which, polysaccharide was removed by aether extraction prior to RNA precipitation. Electrophoresis standard for judging the quality of total RNA was also put forwards in this paper.