GSM9690445: Endometrial biopsy, pre-treatment, rep13; Equus caballus; RNA-Seq
Endometrial biopsy samples were collected from the base of each uterine horn near the uterine body using sterile biopsy forceps. Each sample was immediately divided under sterile conditions, and the portion intended for transcriptomic analysis was preserved in RNAlater solution and stored at −20°C until RNA extraction and subsequent RNA-Seq analysis. Total RNA was extracted from tissue samples using the standard TRI Reagent™ Solution (ThermoFisher Scientific, USA) following the manual homogenisation of samples submerged in TRI Reagent™. For library preparation, 250 ng of total RNA was processed using the QuantSeq 3′ mRNA-Seq Library Prep Kit FWD (Lexogen, Austria), designed to produce a single fragment per transcript from the 3′ end. Library integrity was evaluated with the TapeStation 4150 System (Agilent, USA), and concentrations were measured using the Qubit dsDNA BR assay (Thermo Fisher Scientific, USA). Sequencing was carried out at the OMRF Clinical Genomics Center on an Illumina NovaSeq 6000 platform in paired-end 150 bp mode; however, in line with the QuantSeq 3′ approach, only the first read was used for downstream analysis, while the second read was excluded during preprocessing.