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JoVE Journal Genetics
A Complete Pipeline for Isolating and Sequencing MicroRNAs, and Analyzing Them Using Open Source Tools
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A Complete Pipeline for Isolating and Sequencing MicroRNAs, and Analyzing Them Using Open Source Tools

Article DOI: 10.3791/59901-v • 09:29 min • August 21st, 2019
August 21st, 2019 •
1, 1, 1
1Division of Medical Genetics, University of Washington School of Medicine

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Here, we describe a step-by-step strategy for isolating small RNAs, enriching for microRNAs, and preparing samples for high-throughput sequencing. We then describe how to process sequence reads and align them to microRNAs, using open source tools.

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Isolating Sequencing MicroRNAs Analyzing Open Source Tools Protocol Small RNA Sequencing Normalized Reads Gel Purification Bioinformatics Analysis High Throughput Sequencing Disease Mechanisms Processing Of MicroRNAs Quantification Of Gene Therapy Approaches RNase Free Environment Ice Gel Cutting Steps Three Prime Adaptor Ligation ATP Free 10X T4RNA Ligase Reaction Buffer Polyethylene Glycol Three Prime Linker Thermocycler T4RNA Ligase Two Room Temperature Incubation Polyacyrilamide Gel
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