GSM6394760: mouse islets 20mM glucose 3; Mus musculus; RNA-Seq
RNA concentration was measured using Qubit 3 Fluorometer (ThermoFisher Scientific) and RNA quality was measured using Agilent 2200 TapeStation (Agilent).  RNA sequencing libraries were prepared using the CEL-Seq2 protocol, as published by Hashimshony et al. [Hashimshony et al. Genome Biology (2016) 17:77], with one modification- instead of single-cells as input, 3 ng purified RNA was taken as input for library preparation. The CEL-Seq2 libraries were analyzed for average fragment size using Agilent 2200 TapeStation (Agilent) and concentration was measured using Qubit 3 Fluorometer (Thermo Fisher Scientific). The libraries were sequenced on the Illumina Hiseq 2500 sequencer (Illumina) rapid mode, 15 bases for read 1 and 52 bases for read 2. Demultiplexing was performed in two steps. First, Illumina demultiplexing was performed using bcl2fastq Illumina software with the following parameters: barcode-mismatches =1, minimum-trimmed-read-length = 0, and mask-short-adapter-reads = 0. Second, Cell-seq demultiplexing using the pipeline described in Hashimshony et al. (https://github.com/yanailab/CEL-Seq-pipeline) was executed with the following parameters: min_bc_quality = 15, bc_length = 6, umi_length = 6, and cut_length = 50. RNA measurements, library preparation and sequencing were performed by the Technion Genomics Center, Technion, Israel.