STAR Protocols (Dec 2021)

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

  • Sini Ezer,
  • Masahito Yoshihara,
  • Shintaro Katayama,
  • Carsten Daub,
  • Hannes Lohi,
  • Kaarel Krjutskov,
  • Juha Kere

Journal volume & issue
Vol. 2, no. 4
p. 100995

Abstract

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Summary: 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.

Keywords