草莓不同组织RNA提取方法的改良研究

探讨一种适于草莓不同组织的RNA提取的改良方法。通过增加裂解液震荡强度、离心柱多次吸附、增加去蛋白液温育次数及漂洗次数等步骤对EASYspin RNA提取总RNA操作方法进行改进。结果表明,改良方法提取的草莓根、茎、叶、果实、花中的总RNA,电泳检测都出现清晰的28S和18S两条明显的条带,表明总RNA完整性较好;紫外分光计分析其OD260/OD280值在1.88至2.06之间,OD260/OD230值均大于2.0,RNA浓度含量为310~500μg/mL,表明提取的总RNA纯度好、含量高;基于RNA中mRNA反转录的RT-PCR扩增Actin基因分析结果表明,改良方法提取的草莓不同组织的总RNA质量高。总之,经过电泳、紫外、反转录分析结果证实,改良EASYspin适于草莓果实等不同组织总RNA的提取,是草莓类似果树种类总RNA提取的首选借鉴方法。

Improvement of RNA extraction method from strawberry different tissues

This study explores an improved method for RNA extraction from strawberry different tissues. The improved EASYspin method for RNA extraction includes an increase in lysating-shocking strength, pin-column adsorbing times, removal-of-protein solution incubation times, and elution times. The results showed that two distinct electrophoresis bands of 28 S and 18 S were included in the total RNAs, they were extracted by the improved EASYspin from strawberry different tissues of root, stem, leave, fruit and flowers, which indicated the integrity of total RNAs; the OD260/OD280 value with 1.88-2.06, OD260/OD230 values with 2.0 above, and RNA concentrations with 310-500 μg/mL showed the high purity of total RNAs by UV spectrometer analysis. The results based on RT-PCR analysis using Actin gene showed the high quality cDNAs of the total RNAs. In conclusion, the results from electrophoresis, UV spectrometer, and RT-PCR demonstrated that the improved EASYspin methods are suitable for extraction of total RNA from strawberry different tissues, and are also preferred RNA extraction method for other similar fruit crops to strawberry.