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MicroRNA Expression Analysis: Next-Generation Sequencing

  • Poching Liu
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1783)

Abstract

MicroRNAs are a class of small noncoding RNAs that function as regulators involving in many biological processes. The evaluation of miRNAs and their targets has been aided by miRNA expression profiling studies including multiplex PCR, microarrays, and recent next-generation sequencing tools. Next-generation sequencing has enabled us to profile thousands of genes in a single experiment and overcome the background signal and cross-hybridization issues of microarrays. Next-generation sequencing also allows for the simultaneous confirmation of known miRNAs and discovery of new miRNAs, and significantly reduces costs while providing billions of nucleotide information within a single experiment. Here we describe a detailed procedure of generation of miRNA library for next-generation sequencing to increase the efficiency of adapter ligation and finally construct a more specific cDNA library for sequencing and analyses for miRNA expression profiling.

Key words

MicroRNA miRNA Next-generation sequencing Massive parallel sequencing Expression profiling Microarray RT-PCR Multiplexing 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.DNA Sequencing and Genomics Core—NHLBINational Institute of HealthBethesdaUSA

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