摘要
为建立高效提取几种红景天属植物叶片RNA的方法,试验以采自西藏林芝色季拉山的大花红景天为材料,分析了CTAB法,Trizol法,SDS法,尿素法提取叶片RNA的可行性,并利用Nano Drop 1 000、RTPCR和q RT-PCR等手段分析了该方法获得RNA的质量.结果显示,SDS法和CTAB-异丙醇均可得到完整的RNA条带,但是相比于SDS法,后者所提取的RNA产率较高,质量完整,条带清晰.随后对9种不同产地,不同品种的红景天进行RNA的提取,并用Nano Drop 1 000、RT-PCR分析结果.结果显示,所提取RNA的浓度均在200 ng/μL之上,A260/A280均在2.0-2.2之间,A260/A230均大于1.8.此外利用q RT-PCR获得18S内参基因的标准曲线相关系数为0.983.因此证明所提取的RNA的纯度、完整性和产率较高,蛋白、多糖及多酚类物质去除干净.所以CTAB-异丙醇法可以适用于红景天属叶片RNA的提取,为红景天的分子生物学研究奠定了基础.
In order to establish efficient extraction methods of leaf RNA of several Rhodiola, we used Rhodiola crenulata which were collected from Tibet Sejila Mountain as materials, and analyzed the feasibility of CTAB, Trizol, SDS, urea extraction methods to extract RNA, and used Nano Drop1 000, RTPCR and q RT-PCR to analyze RNA quality. The results showed that both SDS and CTAB-isopropanol method could yieled good results, with the latter being better than the former. Then RNAs were extracted from nine kinds of different origins and species of Rhodiola, and the quality was checked by Nano Drop1000, RT-PCR. The results showed that the concentrations of RNAs were above 200 ng/μL, the values of A260/A280 were between 2.0 and 2.2, and the values of A260/A230 were greater than 1.8.Furthermore, by q RT-PCR, R2 of 18 S standard curve was 0.983. The results presented that the purity,integrity and yield of the RNAs were good, and protein, polysaccharide, polyphenols were removed clearly. So CTAB-isopropanol method could be applied to RNA extraction from leaves of Rhodiola, and this result could contribute to further study of molecular biology in Rhodiola.
出处
《南开大学学报（自然科学版）》
CAS
CSCD
北大核心
2017年第6期48-53,共6页
Acta Scientiarum Naturalium Universitatis Nankaiensis
