摘要
在液氮中研磨小麦幼叶和不同发育时期的种子 ,经含 0 .1%SDS和 0 .1%十二烷基肌氨酸钠 (LDS)的尿素缓冲液裂解后 ,醋酸钠和氯仿沉淀变性蛋白质 ,异丙醇沉淀核酸 ,溶解后经 2 .5mol/LLiCl沉淀总RNA ,洗涤后就可得到高质量的总RNA ,其OD2 60 /OD2 80 为 2 .0 5～ 2 .10 ,2 8S和 18SRNA带清晰 ,叶片总RNA还可得到 2 3S和 16SRNA带 ,产率可达 5mgRNA/ 10g材料。当使用含 1%SDS和 1%LDS的尿素缓冲液裂解材料时 ,则可用于DNA的分离提取 ,其分子大小可达 5 0～ 10 0kb以上。
Wheat leaf and seeds at different development stages had been squashed in liquid nitrogen,then lysised by urea buffer which contains 0.1% SDS and 0.1% LDS,denatured protein had been removed by NaAc and chloroform precipitation,total RNA was further purified by LiCl.The RNA we obtained had sharp bands of 28S and 18S after agarose gel electrophoresis,23S and 16S RNA bands can also be seen clearly in leaf RNA extract,the value of OD 260 /OD 280 of RNA was 2 05～2 10.5mg RNA can been isolated from 10g leaf of wheat.This method can also been used in high molecular weight DNA isolation but the concentration of SDS and LDS must be increased to 1%.
出处
《遗传》
CAS
CSCD
北大核心
2002年第3期337-338,共2页
Hereditas（Beijing)
基金
国家转基因专项 (J99-A -0 37)农杆菌介导小麦转化技术的研究
