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Sci Rep. 2016 Aug 12;6:31584. doi: 10.1038/srep31584.

Globin mRNA reduction for whole-blood transcriptome sequencing.

Author information

1
Department of Biosciences and Nutrition, Karolinska Institutet, Huddinge, Sweden.
2
Competence Centre on Health Technologies, Tartu, Estonia.
3
Molecular Neurology Research Program, University of Helsinki and Folkhälsan Institute of Genetics, Helsinki, Finland.
4
Institute of Molecular and Cell Biology, University of Tartu, Tartu, Estonia.
5
Department of Women's and Children's Health, Karolinska Institutet, Stockholm, Sweden.
6
Department of Reproductive Biology, Estonian University of Life Sciences, Tartu, Estonia.
7
Institute of Biomedicine and Translational Medicine, University of Tartu, Tartu, Estonia.
8
Department of Obstetrics and Gynaecology, University of Tartu, Tartu, Estonia.
9
Department of Obstetrics and Gynecology, Helsinki University Hospital, Helsinki, Finland.

Abstract

The transcriptome analysis of whole-blood RNA by sequencing holds promise for the identification and tracking of biomarkers; however, the high globin mRNA (gmRNA) content of erythrocytes hampers whole-blood and buffy coat analyses. We introduce a novel gmRNA locking assay (GlobinLock, GL) as a robust and simple gmRNA reduction tool to preserve RNA quality, save time and cost. GL consists of a pair of gmRNA-specific oligonucleotides in RNA initial denaturation buffer that is effective immediately after RNA denaturation and adds only ten minutes of incubation to the whole cDNA synthesis procedure when compared to non-blood RNA analysis. We show that GL is fully effective not only for human samples but also for mouse and rat, and so far incompletely studied cow, dog and zebrafish.

PMID:
27515369
PMCID:
PMC4981843
DOI:
10.1038/srep31584
[Indexed for MEDLINE]
Free PMC Article

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