Generation of RNA sequencing libraries for transcriptome analysis of globin-rich tissues of the domestic dog

We have developed a protocol for barcoded cDNA libraries of 48 samples to study gene expression across tissues in the domestic dog, Canis familiaris, by modifying the Single-Cell Tagged Reverse Transcription (STRT) protocol (Islam et al., 2012, 2014). The cDNA reads represent mRNA 5′ ends, enabling...

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Detalles Bibliográficos
Autores principales: Ezer, Sini, Yoshihara, Masahito, Katayama, Shintaro, Daub, Carsten, Lohi, Hannes, Krjutskov, Kaarel, Kere, Juha
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8672047/
https://www.ncbi.nlm.nih.gov/pubmed/34950881
http://dx.doi.org/10.1016/j.xpro.2021.100995
Descripción
Sumario:We have developed a protocol for barcoded cDNA libraries of 48 samples to study gene expression across tissues in the domestic dog, Canis familiaris, by modifying the Single-Cell Tagged Reverse Transcription (STRT) protocol (Islam et al., 2012, 2014). The cDNA reads represent mRNA 5′ ends, enabling the study of transcription start sites (TSS). Our modifications include longer UMIs for molecular counting and Globin-Lock® to deplete globin mRNAs that are abundant in blood and blood-rich tissues dominating all reads.

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