title: GSM5538753: H13NOD20NT; Rattus norvegicus; RNA-Seq
library_construction_protocol: Total RNA was extracted from rat liver tissue samples preserved with an RNA stabilization solution (Thermo scientific, USA). RNA purification was performed with RNeasy Mini Kit® (Qiagen, USA) according to the instructions provided by the manufacturer and quality assessment for the RNA Integrity Number (RIN) was performed using Agilent 2100 Bioanalyzer (Agilent, Palo Alto, CA). Libraries were prepared using the Illumina TruSeq Stranded mRNA kit (Illumina, San Diego, USA), starting with 1 μg of total RNA. mRNAs were purified, fragmented and converted to cDNA with reverse transcriptase. The resulting cDNAs were converted to double stranded cDNAs and subjected to end-repair, A-tailing, and adapter ligation. The constructed libraries were amplified using 15 cycles of PCR. Libraries were quantified by qPCR with the LightCycler 480 real-time PCR instrument (Roche, Basel, Switzerland). Normalized and pooled libraries were sequenced on a NovaSeq 6000 platform with S1 flow cell (200 cycles) (Illumina, San Diego, CA, USA) in 75 bp paired-end.