Equine tissue miRNA profiling

Purpose: Identification of miRNA expression profiles of selected horse tissues. Methods: miRNA-seq analysis was performed on lung, heart and liver samples collected from 4 horses. The miRNA libraries were constructed from total RNA using NEBNext Multiplex Small RNA Library Prep Set for Illumina (New England Biolabs) according to the manufacturer protocol. The quantification of the obtained libraries was performed on a Qubit 2.0 spectrophotometer (Invitrogen, Life Technologies), while a quality control on a TapeStation 2200 instrument (D1000 ScreenTape; Agilent). 75 single-end cycle sequencing was performed on the NextSeq 500/550 platform (Illumina) with the use of NextSeq High Output Reagents v2 (Illumina). Results: The comparison of miRNA profiles between the investigated tissues revealed 185 differentially expressed microRNAs (p adj=0.05) in the heart samples versus the liver samples, 172 DE miRNAs (p adj =0.05) in the lung samples in comparison to the liver samples, and 155 in the lung samples versus the heart samples. Conclusions: Obtained results show varied miRNA expression profiles characteristics for each investigated hore tissue. Overall design: MicroRNA next generation sequencing of selected equine tissues was performed using the Illumina NextSeq 500/550 platform.