title: GSM6133904: High fat diet 2; Mus musculus; RNA-Seq
library_construction_protocol: Cells were washed with cold PBS and RNA was extracted using the Machery Nagel Nucleospin RNA Mini kit for RNA purification (Ref: 740955.50) following manufacturer's instructions. RNA yield was measured using the Qubit Hs RNA assays (Invitrogen. Ref: Q32852) on a QubitTM 3.0 fluorometer (Thermo Fischer Scientific). RNA quality was assessed using the Tapestation 4200 with High Sensitivity RNA screentape (all from Agilent) by the RNA integrity number (RIN) algorithm. Samples with a RIN between 8.5 and 9.4 were used for library preparation. Libraries were prepared using the Ovation® solo RNA-Seq system mouse (Tecan. Ref: 0501), with ribosomal RNA depletion and double indexing following the manufacturer's instructions. Next generation sequencing was performed using the NextSeq 500/550 High Output Kit v2.5 (Illumina. Ref: 20024906) on the NextSeq500 system (Illumina). Runs were carried out over 75 cycles, to obtain 75-bp single reads. Illumina Pipeline software, Casava version 1.9 was used to process the sequencing data.