摘要
针对人参组织多酚、多糖类物质含量较高的特点,比较了改良的异硫氰酸胍法、改良的CTAB法和改良的Trizol法等3种不同的RNA提取方法.3种改良的方法均能从人参组织中提取到总RNA.其中改良的Trizol法能有效地抑制酚类物质和多糖对总RNA提取的影响,能从成熟的叶片中获得高质量、完整性好的总RNA,每克新鲜组织RNA产量在90～120!g之间,电泳分析,28SrRNA亮度约为18SrRNA的2倍,A260/A280介于1.8～2.0之间.用改良的Trizol法分离的RNA,已成功进行了RT-PCR及人参叶cDNA文库构建等研究.
Extraction of high-quality RNA from Panax ginseng tissues is particularly difficult due to high levels of polyphenolics and polysaccharides, or other compounds that bind and/or coprecipitate with RNA. The methods such as guanidinum thioeyanate, CTAB and modified Trizol were applied to isolate high purity and integrity RNA from Panax ginseng. By all of these methods total RNA can be obtained, but only by the modified Trizol method, it extracts integrity and undegraded RNA from mature leaf tissue. Following this efficient procedure, 90 ～ 120μg/g total RNA from leaf tissues can be obtained. The ratio of intensities of the 28S and 18S rRNA bands was 2 ： 1. The absorbance ratios （A260/A280） were 1.8 to 2.0. The RNA is suitable for RT-PCR and eDNA library construction.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2006年第1期95-99,共5页
Progress In Biochemistry and Biophysics
基金
吉林省社会发展计划中药现代化专项重大项目(20011109)~~
关键词
RNA提取
人参
次生物质
RNA extraction, Pnax ginseng, secondary substance
