番木瓜果皮和种子总RNA提取方法的研究  

Total RNA Isolated from Pericarp and Seeds of Carica papaya Linn.

作　　者：王静毅[1] 郑丽珊[1] 冀小蕊[1] 邓柳红[1] 武耀廷[1] 

机构地区：[1]中国热带农业科学院热带生物技术研究所,热带作物生物技术国家重点实验室,海口市571101

出　　处：《热带农业科学》2006年第6期18-20,共3页Chinese Journal of Tropical Agriculture

基　　金：中国热带农业科学院博士后启动基金项目

摘　　要：CTAB法、SDS法和改良的热硼酸法所提取的番木瓜果皮总RNA产量分别是107.8、90.4和70.4μg/g;种子总RNA产量分别是164.0、204.8和111.2μg/g。用TSSE或TE溶液溶解CTAB法中的LiCl过夜沉淀,得到的总RNA完整性好、纯度高,种子总RNA的产量明显提高,达到300μg/g以上。用CTAB法提取的果皮和种子总RNA已成功用于cDNA合成、文库构建和抑制消减杂交(SSH)等后续分子生物学试验。CTAB, SDS and modified hot borate methods were applied to extract high quality RNA from the pericarp and seeds of Carica papaya Linnaeus which contains abundant polyphenols, polysaccharides, proteins, pectins and secondary metabolites. These three methods produced respective total RNA yields of 107.8 μg/g, 90.4 μg/g and 70.4 μg/g from the pericarp, and 164.0 μg/g, 204.8 μg/g and 111.2 μg/g fi＇om the seeds. The improved CTAB method significantly enhanced the yield of high-quality RNA of seeds （more than 300 μg/g）. The extracted RNA from the pericarp and seeds was successfully applied for cDNA synthesis, library construction and suppression subtractive hybridization of subtracted cDNA libraries, which was proved that the isolated total RNA by the improved CTAB method was suitable for the follow-up molecular biological studies.

关 键 词：番木瓜 果皮 种子 总RNA SSH 

分 类 号：Q522[生物学—生物化学]