GSM8207823: Ccr2+MHCII+ DSS2; Mus musculus; RNA-Seq
Total RNA was isolated from sorted cell populations using Trizol protocol as described previously (PMID: 15284119 and PMID: 32276965) RNA-seq libraries were prepared in the Penn State College of Medicine Genome Sciences core (RRID:SCR_021123) using Lexogen's QuantSeq 3' mRNA-Seq V2 Library Prep Kit Forward (FWD) with 12 nucleotide Unique Dual Indices (UDIs) following the manufacturer's instructions for low input RNA library prep. Briefly, 1 ng of total RNA was reverse transcribed using oligo (dT) primers. The second cDNA strand was synthesized by random priming followed by cDNA purification and library amplification using Lexogen's 12 nt Unique Dual Indices and 24 cycles. The libraries were analyzed for size distribution and concentration using BioAnalyzer High Sensitivity DNA kit (Agilent technologies, Santa Clara, CA). Libraries were pooled at equimolar concentrations and sequenced on Novaseq6000 (Illumina) to get approximately 5 million paired end 50 bp reads.