Rodrigues Simone M, Soares Virgínia L F, de Oliveira Tahise M, Gesteira Abelmon S, Otoni Wagner C, Costa Marcio G C
Plant Biology Department, Federal University of Viçosa, Vicosa, Minas Gerais, Brazil.
Mol Biotechnol. 2007 Nov;37(3):220-4. doi: 10.1007/s12033-007-0070-9. Epub 2007 Aug 8.
The tropical plant Bixa orellana L. (annatto) produces an array of natural products, including the pigment bixin used in the food and cosmetics industries. In order to understand the biochemical and molecular basis of the biosynthesis of these natural products, a reliable method for isolating high yields of high-quality RNA is required. Here we described a successful and reproducible method for isolation and purification of high-quantity and high-quality RNA from different tissues of annatto. This protocol overcomes the usual problems associated with large amounts of polyphenols, polysaccharides, pigments, and other secondary metabolites that are not easily removed by conventional extraction procedures. Furthermore, the proposed protocol can be easily carried out in any laboratory and it could also be extended to isolate RNA from other plant species showing similar abundance of compounds that interfere with RNA extractions. The yield and quality of the RNA were monitored by spectrophotometric analysis, separation on agarose gel, Reverse Transcription-Polymerase Chain Reaction (RT-PCR), and construction of a cDNA library.
热带植物红木（Bixa orellana L.）能产生一系列天然产物，包括食品和化妆品工业中使用的色素胭脂树橙素。为了了解这些天然产物生物合成的生化和分子基础，需要一种可靠的方法来分离高产率的高质量RNA。在此，我们描述了一种从红木不同组织中分离和纯化高产量、高质量RNA的成功且可重复的方法。该方案克服了与大量多酚、多糖、色素和其他次生代谢产物相关的常见问题，这些物质不易通过传统提取程序去除。此外，所提出的方案可以在任何实验室轻松实施，并且还可以扩展到从其他含有类似干扰RNA提取化合物的植物物种中分离RNA。通过分光光度分析、琼脂糖凝胶分离、逆转录-聚合酶链反应（RT-PCR）以及构建cDNA文库来监测RNA的产量和质量。
