Messenger RNA capture sequencing of extracellular RNA from human biofluids using a comprehensive set of spike-in controls

Autor: 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
Jazyk: angličtina
Rok vydání: 2021
Předmět:
Zdroj: STAR Protocols, Vol 2, Iss 2, Pp 100475- (2021)
Druh dokumentu: article
ISSN: 2666-1667
DOI: 10.1016/j.xpro.2021.100475
Popis: Summary: 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).
Databáze: Directory of Open Access Journals