Globin mRNA reduction for whole-blood transcriptome sequencing.
Sci Rep
; 6: 31584, 2016 08 12.
Article
em En
| MEDLINE
| ID: mdl-27515369
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.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Células Sanguíneas
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Globinas
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RNA Mensageiro
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Sequenciamento de Nucleotídeos em Larga Escala
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Transcriptoma
Limite:
Female
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Humans
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Male
Idioma:
En
Ano de publicação:
2016
Tipo de documento:
Article