Of the 358 endometrial samples, 290 had Illumina Human HT-12 v4.0 performed and 266 underwent RNA-seq (198 samples had both techniques performed). Total RNA was isolated from endometrial samples using the Allprep DNA/RNA Mini Kit (Qiagen, CA) as per the manufacturer’s instructions29 (link). Briefly, RNA quality was checked using a Bioanalyzer 2100 (Agilent Technologies, CA) and RNA concentration was measured using a NanoDropND-6000 (Thermo Fisher Scientific, USA). All samples were high quality with an RNA integrity number greater than 8. Expression profiles in endometrial tissue were generated by hybridizing 750 ng of cRNA to Illumina Human HT-12 v4.0 Beadchips.
RNA samples were treated with Turbo DNA-free kit (Thermo Fisher Scientific, USA) prior to RNA-seq library generation19 (link). Stranded RNA-seq libraries were prepared using the Illumina TruSeq Stranded Total RNA Gold protocol which includes ribosomal depletion (Illumina, USA).
RNA samples were treated with Turbo DNA-free kit (Thermo Fisher Scientific, USA) prior to RNA-seq library generation19 (link). Stranded RNA-seq libraries were prepared using the Illumina TruSeq Stranded Total RNA Gold protocol which includes ribosomal depletion (Illumina, USA).