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Updated: Nov 7, 2025
Sequencing of mRNA from Whole Blood using Nanopore Sequencing
Published on: June 3, 2019
Messenger RNA capture sequencing of extracellular RNA from human biofluids using a comprehensive set of spike-in
Eva Hulstaert1,2,3, Anneleen Decock1,2, Annelien Morlion1,2
1Center for Medical Genetics, Department of Biomolecular Medicine, OncoRNALab, Ghent University, C. Heymanslaan 10, 9000 Ghent, Belgium.
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).
