GSM3931526: Q109-CONTROL; Equus caballus; RNA-Seq
Total cellular RNA was extracted from endometrial samples using a RNeasy Mini Kit (Qiagen, Gaithersburg, MD). A total of 1 μg of RNA was treated with DNase I (Ambion Inc., Austin, TX) for 30 minutes at 37℃ to remove genomic DNA according to manufacturer's instructions. Endometrial samples were processed for RNA extraction using a RNeasy Mini Kit (Qiagen, Gaithersburg, MD) following the manufacturer's recommendations. After extraction, RNA concentration and quality were analyzed using a Nanodrop 2000 spectrophotometer Thermo Fisher Scientific) and a Bioanalyzer® (Agilent, Santa Clara, CA). Samples had a 260/280 ratio > 2.0, 28S:18S rRNA ratios >1.8 and RNA integrity number (RIN) was 9.78  0.06 (mean  SEM). A TruSeq Stranded mRNA library prep kit (Illumina, San Diego, CA) was used to prepare libraries for mRNA sequencing. Libraries were loaded onto an Agilent DNA 1000 chip and validated on an Agilent 2100 Bioanalyzer (Agilent). Quantitation was performed with the Illumina Library Quantification Kit, ABI Prism qPCR Mix from Kapa Biosystems. Libraries were run on a NextSeq 500 v2 (Illumina) 300cycles High Output kit in a 2x150 base pairs with paired-end reads.