Methods for Isolation of Total RNA to Recover miRNAs and Other Small RNAs from Diverse Species
Methods for Isolation of Total RNA to Recover miRNAs and Other Small RNAs from Diverse Species
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DOI：
10.1007/978-1-60327-005-2_3
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发表时间：
2010-01-01
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影响因子：
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通讯作者：
Green, Pamela J.
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文献类型：
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作者：
Accerbi, Monica;Schmidt, Skye A.;Green, Pamela J.
对于20-25个核苷酸（nt）大小范围内的mirna和其他小RNA的实验分析，第一步也是最重要的一步是分离高质量的总RNA。因为RNA降解产物会掩盖或稀释真正的mirna的存在，所以在选择一种方法时，重要的是它能有效地从组织中提取RNA，同时防止高分子量和低分子量RNA的降解。此外，多酚、多糖和次生代谢物的存在可能使核酸不溶，并阻碍mirna的恢复。最后，也是最重要的，所选择的方法必须能够保留小RNA成分。在本章中，我们将提出一套总RNA分离方法，可以最大限度地回收高质量的RNA，用于大量植物物种和组织类型的miRNA分析。
For the experimental analysis of miRNAs and other small RNAs in the 20-25 nucleotide (nt) size range, the first and most important step is the isolation of high-quality total RNA. Because RNA degradation products call mask or dilute the presence of true miRNAs, it is important when choosing a method that it efficiently extracts RNA from tissues in a manner that prevents degradation of RNA of both high and low molecular weight. In addition, the presence of polyphenols, polysaccharides, and secondary metabolites may render nucleic acids insoluble, and hinder the recovery of the miRNAs. Finally, and most importantly, the method chosen must be capable of retaining the small RNA component. In this chapter, we will present a set of total RNA isolation methods that can be used to maximize the recovery of high-quality RNA to be used in miRNA analysis for a large number of plant species and tissue types.