A general method for high-quality RNA isolation from metabolite-rich fruits
A general method for high-quality RNA isolation from metabolite-rich fruits
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DOI：
10.1016/j.sajb.2012.08.004
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发表时间：
2012-11-01
影响因子：
3.1
通讯作者：
Defilippi, B. G.
中科院分区：
文献类型：
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作者：
Gudenschwager, O.;Gonzalez-Aguero, M.;Defilippi, B. G.
从水果组织中分离高质量的 RNA 通常很困难，因为水果组织中含有丰富的多糖、酚类物质和其他代谢物，这些代谢物会对分离的 RNA 的质量和产量产生负面影响。在这里，我们描述了对现有方法的一些修改，该方法可用于从含有大量代谢物的几种不同水果的果肉中分离 RNA。与其他测试的提取方案相比，该方法已成功用于从所有测试物种的两个发育阶段提取大量高质量的总 RNA，并通过分光光度法和变性琼脂糖凝胶电泳进行测定。该 RNA 还适用于逆转录聚合酶链式反应和实时定量 RT-PCR 等下游应用。这里描述的方法可能用于研究多种水果组织发育和成熟过程中的基因表达，特别是在其他方法无法产生合适 RNA 的情况下。 (C) 2012 萨博。由 Elsevier B.V. 出版。保留所有权利。
The isolation of high-quality RNA from fruit tissues is often difficult due to the abundance of polysaccharides, phenolics and other metabolites that can negatively affect both the quality and yield of isolated RNA. Here, we describe several modifications to an existing method that can be used to isolate RNA from the pulp of several different fruits that contain large amounts of metabolites. This method, in contrast to other extraction protocols tested, was successfully used to extract large quantities of high-quality total RNA from two developmental stages in all species tested, as assayed by spectrophotometry and electrophoresis on denaturing agarose gels. This RNA was also suitable for use in downstream applications such as reverse transcription-polymerase chain reaction and real-time quantitative RT-PCR. The method described here could likely be used for studying gene expression during the development and ripening of a wide variety of fruit tissues, especially in cases where other methods fail to yield suitable RNA. (C) 2012 SAAB. Published by Elsevier B.V. All rights reserved.