Globin mRNA reduction for whole-blood transcriptome sequencing

Date
2016Author
Krjutškov, Kaarel
Koel, Mariann
Roost, Anne Mari
Katayama, Shintaro
Einarsdottir, Elisabet
Jouhilahti, Eeva-Mari
Söderhäll, Cilla
Jaakma, Ülle
Plaas, Mario
Vesterlund, Liselotte
Lohi, Hannes
Salumets, Andres
Kere, Juha
Metadata
Show full item recordAbstract
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.