Guangdong Provincial Key Lab of Biotechnology for Plant Development, College of Life Sciences, South China Normal University, Guangzhou, People's Republic of China.
Prep Biochem Biotechnol. 2012;42(1):87-96. doi: 10.1080/10826068.2011.566297.
To isolate high-quality total RNA from Fallopia multiflora tuberous roots is difficult because of the presence of high levels of carbohydrates, phenolics, and other secondary metabolites. Since several procedures specialized for RNA isolation from polysaccharides and phenols rich tissues have resulted in poor yields, in this study, we developed a modified protocol that was derived from the traditional CTAB method. The protocol was able to produce high-quality and intact RNA from the tuberous roots of F. multiflora. The yield of total RNA was more than 0.15 mg/g fresh weight, with an A260/A280 ratio of 1.9-2.0. The obtained RNA was of sufficient quality and suitable for downstream application such as reverse-transcription polymerase chain reaction (RT-PCR), Northern hybridization, and cDNA library construction. The protocol may also have wider applicability for total RNA isolation from other plant species with tuberous roots.
从虎杖块根中分离高质量的总 RNA 较为困难，因为其含有高水平的碳水化合物、酚类化合物和其他次生代谢物。由于几种专门针对富含多糖和酚类化合物组织的 RNA 分离程序导致产量较低，因此在本研究中，我们开发了一种源自传统 CTAB 方法的改良方案。该方案能够从虎杖块根中获得高质量和完整的 RNA。总 RNA 的产量超过 0.15mg/g 鲜重，A260/A280 比值为 1.9-2.0。所获得的 RNA 质量足够好，适用于下游应用，如逆转录聚合酶链反应 (RT-PCR)、Northern 杂交和 cDNA 文库构建。该方案可能也适用于从其他具有块根的植物物种中分离总 RNA。
