title: GSM5824552: BMI Epididymis RNAseq rep1; Sus scrofa; RNA-Seq
library_construction_protocol: Total RNA was extracted from five mixed testicles samples and five mixed epididymal samples using TRIzol (Thermo Fisher Scientific, Invitrogen, Catalog #15596018) following the protocol provided by the manufacturer. The RIN (RNA integrity number) value of RNA samples was determined by an Agilent 2100 Bioanalyzer and 1% agarose gel electrophoresis. The quality control step ensures that RIN values are above 7.5 and 28S/18S ratios are greater than or equal to 1.5. The purity and concentration of RNAs were detected with a Nanodrop microspectrophotometer (Thermo Fisher Scientific). Oligo(dT) magnetic beads were used to enrich RNAs with poly-A tails and a SMARTer cDNA synthesis kit (Takara, Clontech, Catalog #634925) was used to generate full-length cDNA molecules. For Illumina RNAseq, the cDNA fragments were purified with a QiaQuick PCR extraction kit (Qiagen, Venlo, The Netherlands, Catalog # 28106), followed by end repair, base addition, and ligation to sequencing adapters. The ligation products were further size selected by agarose gel electrophoresis and amplified by PCR.