GSM3108318: 4410-FFB-0003_miRNAseq; Sus scrofa domesticus; miRNA-Seq
Total RNA was isolated using Rneasy kit (Qiagen) following the manufacturer's protocol.  RNA quality was assessed using Agilent Bioanalyzer and Qbit Fluorometer. Samples with an RNA integrity number (RIN) ≥ 7.5 were used for library preparation for RNA sequencing RNAseq) and miRNA sequencing (miRNAseq). Briefly, total RNA (500 ng) from each endometrial sample was used for mRNA and miRNA library preparation. For mRNAseq, polyadenylated (polyA) RNAs were isolated using NEBNext Magnetic Oligo d(T)25 Beads (New England BioLabs, Inc.). After polyA selection, RNA was fragmented and primed for first-strand synthesis using the NEBNext First Strand Synthesis Module, followed by second-strand synthesis using the NEBNext Second Strand Synthesis Module. Samples followed standard library preparation protocol using NEBNext DNA Library Prep Master Mix Set for Illumina. For miRNAseq, the NEBNext Small RNA Library Prep Set for Illumina (New England Biolabs Inc., Ipswich, MA, USA) was followed according to the manufacturer's protocol. Final mRNA and miRNA library quantities were assessed using a Qubit® 2.0 Fluorometer and library quality was determined using a DNA High Sense chip on Agilent 2100 Bioanalyzer. Further library quantification was completed using the qPCR-based KAPA Biosystem Library Quantification kit (Kapa Biosystems Inc., Woburn, MA, USA). For mRNAseq, each library was diluted to a final concentration of 12.5 nM. For miRNAseq, individual sample libraries were diluted to a final concentration of 1.25 nM. Equimolar amounts of individual barcoded  samples from each group were pooled prior to sequencing. Paired-end (50bp) RNAseq at over 25 million reads per individual barcoded sample  was performed using a HiSeq 2500 (Illumina, Inc., San Diego, CA, USA). miRNAseq was conducted using an Illumina HiSeq 2500 instrument (Illumina Inc., San Diego, CA, USA) at 50bp Single End condition generating approximately 15 million reads per sample.
GEO Accession
GSM3108318