title: GSM6436911: IngHFD, WT, Mouse4, Rep2; Mus musculus; RNA-Seq
library_construction_protocol: Tissues were harvested and immediately snap frozen in liquid nitrogen and stored at -80°C. For RNA isolation, 30-50 mg of tissue was placed in Lysing Matrix D tubes and homogenized in 800 µl TRI Reagent (T9424, Sigma) using the Fastprep-24 Homogenizer for 30 s at 4-6 m/s (MP Biomedical). Homogenate was transferred to an RNase free tube and 200 µl chloroform (Sigma) added. The samples were vortexed and centrifuged at 13,000 rpm for 15 min at 4°C. The upper phase was then transferred to an RNase free tube and mixed with an equal volume of 70% ethanol before loading onto RNA isolation spin columns. RNA was extracted using a RNeasy Mini Kit (74106, Qiagen) isolation kit following the manufacturer's instructions. cDNA libraries were made using Illumina TruSeq RNA sample kits and sequencing was performed on Illumina NovaSeq 6000 with paired-end 150 bp reads (Novogene, Cambridge, UK). Raw reads all passed quality control for Qscore, error rate distribution, and AT/GC distribution.