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Messenger RNA Capture Sequencing of Extracellular RNA from Human Biofluids Using a Comprehensive Set of Spike-in Controls

Overview
Journal STAR Protoc
Publisher Cell Press
Specialties Biology
Biomedical Engineering
Science
Date 2021 May 3
PMID 33937877
Citations 5
Authors
Eva Hulstaert
Anneleen Decock
Annelien Morlion
Celine Everaert
Kimberly Verniers
Justine Nuytens
Nele Nijs
Gary P Schroth
Scott Kuersten
Stephen M Gross
Pieter Mestdagh
Jo Vandesompele
Affiliations
Soon will be listed here.
Abstract

Comprehensive transcriptome analysis of extracellular RNA (exRNA) purified from human biofluids is challenging because of the low RNA concentration and compromised RNA integrity. Here, we describe an optimized workflow to (1) isolate exRNA from different types of biofluids and (2) to prepare messenger RNA (mRNA)-enriched sequencing libraries using complementary hybridization probes. Importantly, the workflow includes 2 sets of synthetic spike-in RNA molecules as processing controls for RNA purification and sequencing library preparation and as an alternative data normalization strategy. For complete details on the use and execution of this protocol, please refer to Hulstaert et al. (2020).

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