RNA-Seq of Mus musculus
RNA quality is checked on the 2100 Bioanalyzer (Agilent, USA) using Agilent RNA 6000 Nano Reagents. Only high-quality RNA was sent for sequencing, specifically with a RNA Integrity value (RIN) of more than 7. Livers that were visually affected by cirrhosis (and not reflective of the sample size) were excluded, as well as livers that expressed extreme glucose values during the GTT. Then, seven liver RNA samples from each of the six treatment groups were selected at random and sent to Macrogen (South Korea) for sequencing. Macrogen constructed cDNA and ligated it to Illumina adapters at the 3 and 5 ends using a TruSeq library preparation kit with PolyA pulldown. cDNA libraries were fractionated on agarose gel, and 150 bp fragments were excised and PCR (polymerase chain reaction) amplified.  For cluster generation, the library was loaded into a flow cell where fragments were captured onto a lawn of surface-bound oligos complementary to the library adapters. Each fragment was then amplified into distinct clonal clusters by bridge amplification in the Illumina NovaSeq for a 40 million read depth.
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