Characterization of Plant Small RNAs by Next Generation Sequencing

Q1 Agricultural and Biological Sciences Current protocols in plant biology Pub Date : 2017-03-03 DOI:10.1002/cppb.20043

Sandra M. Mathioni, Atul Kakrana, Blake C. Meyers

引用次数: 28

Abstract

Plant small RNAs are ∼20 to 24 nucleotide noncoding RNAs that typically have repressive regulatory roles in gene expression, functioning at the transcriptional or post-transcriptional level. This influence on regulation of developmental and physiological processes has direct effects on phenotype. High-throughput sequencing technologies have enabled the sequencing of millions of small RNAs. Along with decreased sequencing costs, recent improvements in small RNA library construction have facilitated the ability to use minimal amounts of input RNA for analysis. This unit describes steps to isolate total RNA from limited amounts of plant tissue to construct small RNA libraries and perform small RNA data processing. © 2017 by John Wiley & Sons, Inc.

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植物小rna的下一代测序研究

植物小rna是约20至24个核苷酸的非编码rna，通常在基因表达中具有抑制调节作用，在转录或转录后水平起作用。这种对发育和生理过程调节的影响对表型有直接影响。高通量测序技术已经能够对数百万个小rna进行测序。随着测序成本的降低，最近小RNA文库建设的改进促进了使用最少量输入RNA进行分析的能力。本单元描述了从有限数量的植物组织中分离总RNA的步骤，以构建小RNA文库并进行小RNA数据处理。©2017 by John Wiley &儿子,Inc。

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Current protocols in plant biology Agricultural and Biological Sciences-Plant Science

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期刊介绍： Sound and reproducible laboratory methods are the foundation of scientific discovery. Yet nuances that are critical for an experiment''s success are not captured in the primary literature but exist only as part of a lab''s oral tradition. Current Protocols in Plant Biology provides reproducible step-by-step instructions for protocols relevant to plant research. Furthermore, Current Protocols content is thoughtfully organized by topic for optimal usage and to maximize contextual knowledge. Quarterly issues allow Current Protocols in Plant Biology to constantly evolve to keep pace with the newest discoveries and developments. Current Protocols in Plant Biology is the comprehensive source for protocols in the multidisciplinary field of plant biology, providing an extensive range of protocols from basic to cutting edge. Coverage includes: Extraction and analysis of DNA, RNA, proteins Chromosome analysis Transcriptional analysis Protein expression Metabolites Plant enzymology Epigenetics Plant genetic transformation Mutagenesis Arabidopsis, Maize, Poplar, Rice, and Soybean, and more.