title: GSM8655258: Group 3 Humanized Mgat2 mice fed a HFD. RNASeq performed at the end of the study #11; Mus musculus; RNA-Seq
library_name: GSM8655258
library_construction_protocol: Total RNA was isolated from liver tissue with the RNeasy® Mini kit (Qiagen, Germantown, MD). Liver tissue was homogenized in in RLT lysis buffer using a Bullet Blender® 24 Gold (Next Advance, Raymertown, NY).  200-proof (100%) ethyl alcohol (Pharmco by Greenfield Global, Toronto, Canada) was added and homogenized samples were applied to a RNeasy column.   The column was washed with RW1 buffer and subjected to DNase digestion with a RNase-Free DNase Set (Qiagen, Germantown, MD).  RNA was washed again with RNeasy RW1 and RPE buffers. Total RNA was isolated in RNase-free water, quantified using a NanoDrop® ND-1000 spectrophotometer (ThermoFisher Scientific, Waltham, MA), and stored at -80°C. Cell extracts from 15 mg of liver tissue was used for RNASeq analysis.  RNA quantity was measured using Qubit® (Thermo Scientific, Eugene, OR, United States).  A 4150 TapeStation System was used to determine RNA quality.  An RNA integrity (RIN) score > 7 (10 is maximal) was required for going forward with sequencing.  RNA libraries were prepared using Illumina Stranded Total RNA Prep and the Ribo-Zero™ Plus” kit (Illumina, San Diego, CA, United States).