Plant Developmental Biology
Plant Developmental Biology
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DOI：
10.1007/978-1-60761-765-5
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
L. Hennig;C. Köhler
中科院分区：
文献类型：
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作者：
L. Hennig;C. Köhler
在植物中，基因表达在几个水平上受到调控，最近发现的调控层之一涉及短RNA。短RNA通过几种途径产生，并靶向mRNA或基因组DNA。不同种类的短RNA具有略微不同的大小，并且检测它们的积累是验证和研究非编码短RNA的重要步骤。北方印迹通常用于检测短RNA，因为它提供了关于所分析的短RNA的量和大小的信息。当研究短RNA时，选择正确的RNA提取方案是至关重要的，因为几种常规使用的商业RNA提取试剂盒不产生任何短RNA。本章介绍了优化的RNA提取方法，这给短RNA的良好产量，和分离，转移和杂交协议，研究短RNA的积累。
Gene expression is regulated at several levels in plants, and one of the most recently discovered regulatory layers involve short RNAs. Short RNAs are produced through several pathways and target either mRNAs or genomic DNA. Different classes of short RNAs have slightly different sizes and detection of their accumulation is an important step in validating and studying non-coding short RNAs. Northern blotting is routinely used to detect short RNAs because it gives information about both the amount and size of the analysed short RNAs. Choice of the right RNA extraction protocol is crucial when short RNAs are being studied, because several routinely used commercial RNA extraction kits do not yield any short RNAs. This chapter describes optimised RNA extraction methods, which give good yields of short RNAs, and separation, transfer and hybridisation protocols to study the accumulation of short RNAs.