title: GSM5732376: HFD-O304 islet rep2 - RNA-seq; Mus musculus; RNA-Seq
library_construction_protocol: Mice were killed by cervical dislocation and the pancreata were perfused with 2.5 ml cold collagenase (without Mg2+/Ca2+, GIBCO #14180). The pancreata were then dissected out and incubated at 37 °C for 10–15 min. Collagenase activity was stopped by adding 25 ml of Hanks' solution (with Mg2+/Ca2+, GIBCO #14060) and vigorous shaking released the islets from the rest of the pancreatic tissue. Islets were rinsed in icecold Hanks' three times and then picked manually under a stereo microscope. Isolated islets from 5 mice were pooled and 150 islets were used for extraction of total RNA islets using RNeasy Micro Kit (Qiagen, #74004). RNA-seq libraries were prepared with the Illumina TruSeq RNA Library Kit according to the manufacturer's instructions.