摘要
采用常规的方法提取富含油脂的植物组织总RNA常难以取得满意的结果.对林莎两次裂解法进行了改良,即将SDS的浓度降低至1%,所用酸酚的pH值降低至4.0,去掉提取液II处理步骤,获得的总RNA效果较好:得率高,RNA产量可达74.7μg(100mg)-1;没有DNA污染,A260nm/A280nm值为2.026,A260nm/A230nm值为2.082,说明蛋白质及其它污染非常低,通过RT-PCR成功地扩增出了麻疯树curcin基因长为927bp的特异条带.相比林莎的方法,改良SDS酸酚法具有经济、简便、适用性广的特点.
The total RNA extraction from plant tissues richin lipid by conventional methods often cannot meet the requirement of downstream experiments. This problem was overcomed by modifying Lin＇s twice-extraction method： Adjusting the concentration of SDS and pH value of phenol to 1% and 4.0 respectively, and removing the solution II treatment. The yield was high and reached 74.7 μg （100 mg）^-1 （fresh weight）. There was no genomic DNA contamination in the process. The value of A260 nm/A280 nm was 2.026 and that of A260 nm/A230 nm was 2.082, meaning that protein and other contaminations were low. A 927 bp fragment of Jatropha curcas＇ curcin gene was successfully amplified from the obtained total RNA. Compared to Lin＇s method, this improved SDS acid phenol method is economical, convenient and has a wide applicability. Fig 5, Tab 1, Ref 13
出处
《应用与环境生物学报》
CAS
CSCD
北大核心
2009年第2期284-287,共4页
Chinese Journal of Applied and Environmental Biology
基金
国家自然科学基金项目(No.30670204)
科技部国际合作项目(No.2006DFB63400)
教育部博士点基金项目(No.20060610015)
国家"十一五"科技支撑项目(No.2006BAD07A04)资助~~
关键词
总RNA
麻疯树
油脂
SDS酸酚法
胚乳
total RNA
Jatropha curcas
oil
SDS acid phenol method
endosperm
