GSM8186600: pTh17, shClns1a, 24hr stimulated, Rep 3; Mus musculus; RNA-Seq
Transduced cells were enriched by FACS sorting. Total RNA was extracted using the RNeasy Micro RNA Isolation Kit (Qiagen). RNA quality and quantity were analyzed on a Nanochip cartridge run on the Agilent 2100 Bioanalyzer system. RNAseq libraries were prepared using the Illumina TruSeq stranded total RNA kit (Illumina), starting from 100 ng of DNAse I (Qiagen) treated total RNA, following the manufacturer's protocol with 15 PCR cycles. The amplified libraries were purified using AMPure beads (Beckman Coulter), quantified by Qubit 2.0 fluorometer (Life Technologies), and visualized using the high sensitivity DNA screentape on the Agilent Tapestation 2200 system. The libraries were pooled equimolarly, and loaded onto a P2 100 cycle flow cell (run as paired end 50) and run on the Illumina Nextseq 2000 system.