title: GSM9378235: Sample 1, PB, with multifocal lesions; Bos taurus; RNA-Seq
library_construction_protocol: RNA was extracted from PB, JE, and JELN of the 15 cows included in the study. PB samples were collected from the coccygeal vein of cows using PAXgene Blood RNA tubes (2.5 ml) (Qiagen, Hilden, Germany). Total RNA was extracted with the PAXgene blood RNA kit according to manufacturer's instructions (Qiagen, Hilden, Germany). For RNA isolation from gut samples, 150–200 mg of JE and JELN were harvested and immediately submerged in 2 ml of RNA later (Sigma, St. Louis, MO). Purification of tissue RNA was performed using the RNeasy Mini Kit according to the manufacturer's instructions (Qiagen, Hilden, Germany). Residual DNA was removed by digestion with RNase-free DNase I Amplification Grade (Invitrogen, Spain). Each RNA library was prepared with approximately 250 ng of RNA using the Illumina NEBNext® Ultra Directional RNA library preparation kit (Illumina, San Diego, CA, US). The RNA-Seq libraries' quality was assessed on the Agilent Bioanalizer using a highly sensitive DNA chip to confirm that the insert size was between 188-274 bp for all individual libraries. All RNA-Seq libraries were quantified using a Qubit® Fluorometer and doubled stranded DNA high Sensitivity Assay Kit (Invitrogen, Spain). Libraries were sequenced by single-end sequencing in 1x100 format on the Illumina NextSeq sequencer at the Genomic Unit of the Scientific Park of Madrid, Spain.