title: GSM7077921: C37 104885-001-002; Mus musculus; RNA-Seq
library_construction_protocol: For next generation sequencing the RNA-Bee total-RNA isolation kit (Bio-Connect, Huissen, The Netherlands) was used to isolate total RNA from individual liver samples (lobus sinister lateralis) and RNA integrity was determined with the RNA 6000 Nano Lab-on-a-Chip kit and bioanalyzer 2100 (Agilent Technologies, Amstelveen, The Netherlands). Total RNA's were processed into tagged random sequence libraries. Next Generation Sequencing was performed at Genome Scan (Leiden, the Netherlands). For this, strand-specific cDNA libraries were generated from total-RNA using oligo-dT magnetic beads, mRNA fragmentation, NEBNext Ultra Directional RNA Library Prep Kit from Illumina, NEB #E7760S/L resulting in 300–500 bp amplified libraries/sample. Libraries were multiplexed, clustered, and sequenced on a NovaSeq6000 v1.5 system (Illumina, New England Biolabs, Inc. Ipswich, MA, USA), with a paired-end, 150 bp sequencing protocol, 20-35 million reads per sample and indexing. Clustering and DNA sequencing using the NovaSeq6000 v1.5 was performed according to manufacturer's protocols. A concentration of 1.1 nM of DNA was used yielding paired end reads (2x 150bp). NovaSeq control software NCS v1.7 was used. Image analysis, base calling, and quality check was performed with the Illumina data analysis pipeline RTA3.4.4 and Bcl2fastq v2.20.