Department of Plant Science, School of Biological Sciences, Central University of Kerala, Kasaragod, Kerala, 671316, India.
Biotechniques. 2023 Jun;74(6):302-316. doi: 10.2144/btn-2022-0078. Epub 2023 Jun 8.
Secondary metabolites in mangroves often interfere with RNA extraction yielding poor concentration and quality, which is unsuitable for downstream applications. As existing protocols yielded low-quality RNA from root tissues of (L.) Druce and Lam., an optimized method was developed for improving the quality and yield of RNA. Compared with three other methods, this optimized protocol gave better RNA yield and purity for both species. The absorbance ratios were ≥1.9 for A260/280 and A260/230, while RNA integrity number values ranged from 7.5 to 9.6. Results show that our modified method is efficient in obtaining high-quality RNA from mangrove roots and is suitable for downstream experiments such as cDNA synthesis, real-time quantitative PCR and next-generation sequencing.
红树林中的次生代谢产物常干扰 RNA 提取，导致浓度和质量较差，不适合下游应用。由于现有方案从 (L.) Druce 和 Lam. 的根组织中提取的 RNA 质量较差，因此开发了一种优化方法来提高 RNA 的质量和产量。与其他三种方法相比，该优化方案可提高两种物种的 RNA 产量和纯度。A260/280 和 A260/230 的吸光度比值均≥1.9，而 RNA 完整性数（RIN）值范围为 7.5 至 9.6。结果表明，我们的改良方法可有效地从红树林根部获得高质量的 RNA，适用于 cDNA 合成、实时定量 PCR 和下一代测序等下游实验。
