GSM4779221: T15_IL4_1; Mus musculus; RNA-Seq
Total RNA was isolated from primary microglia using the miRNeasy 96 kit (Qiagen). Libraries were created using commercially available kits according to the manufacturer's instructions, where first strand cDNA synthesis was poly(A)-primed, and then fragmented (SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing, Takara). After fragmentation the library was prepared with the SMARTer ThruPLEX DNA-Seq Kit (Takara). During PCR amplification 10-12 cycles were used. After PCR amplification, the resulting fragments were purified, pooled, quantified and used for cluster generation. For sequencing, pooled libraries were loaded on the cBot (Illumina) and cluster generation was performed according the manufacturer's instructions. Paired-end sequencing using 100 bp read length (multiplex 12 samples per lane - 28M reads) was performed on a Illumina HiSeq 2500 (HiSeq Control Software 2.2.58) using HiSeq Flow Cell v4 and TruSeq SBS Kit v4.