title: GSM4086369: Input_HFD4; Mus musculus; RNA-Seq
library_construction_protocol: 10-week-old male C57/BL6N mice were put either on a control diet or a high fat diet for 3 days. Afterwards, mice were sacrificed by cervical dislocation. The hypothalamus was rapidly dissected using a stainless steel brain matrix (World Precision Instruments) and immediately frozen in liquid nitrogen. Hypothalamic tissues were pooled (8 per IP, 4 IPs per condition) Due to low amount of input material, pre-amplification using the Ovation RNASeq System V2 was performed. Total RNA was used for first strand cDNA synthesis, using both poly(T) and random primers, followed by second strand synthesis and isothermal strand-displacement amplification. For library preparation, the Illumina Nextera XT DNA sample preparation protocol was used, with 1 ng cDNA input.