qRT-PCR of Small RNAs
qRT-PCR of Small RNAs
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DOI：
10.1007/978-1-60761-646-7_10
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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：
Hellens, Roger P.
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文献类型：
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作者：
Varkonyi-Gasic, Erika;Hellens, Roger P.
植物小RNA是一类19- 25个核苷酸（nt）的RNA分子，对基因组稳定、发育和分化、疾病、细胞通讯、信号传导以及对生物和非生物胁迫的适应性反应至关重要。小RNA包括两大类RNA：短干扰RNA （sirna）和微RNA （miRNAs）。高效、可靠地检测和定量小RNA表达已成为了解其在特定细胞和组织中的作用的重要步骤。在这里，我们提供了通过茎环RT-PCR检测mirna的方案。该方法能够快速可靠地从植物组织中提取20 pg的总RNA进行miRNA表达谱分析，适用于高通量miRNA表达分析。此外，该方法可用于检测其他类型的小rna，前提是序列已知且其GC含量与mirna特异性相似。
Plant small RNAs are a class of 19- to 25-nucleotide (nt) RNA molecules that are essential for genome stability, development and differentiation, disease, cellular communication, signaling, and adaptive responses to biotic and abiotic stress. Small RNAs comprise two major RNA classes, short interfering RNAs (siRNAs) and microRNAs (miRNAs). Efficient and reliable detection and quantification of small RNA expression has become an essential step in understanding their roles in specific cells and tissues. Here we provide protocols for the detection of miRNAs by stem-loop RT-PCR. This method enables fast and reliable miRNA expression profiling from as little as 20 pg of total RNA extracted from plant tissue and is suitable for high-throughput miRNA expression analysis. In addition, this method can be used to detect other classes of small RNAs, provided the sequence is known and their GC contents are similar to those specific for miRNAs.