GSM8862566: Gonodal WAT macrophages, control sample 12, LysMCretdT>Fthfl/fl bone marrow chimera; Mus musculus; RNA-Seq
Viable, CD45+, CD11b+, dTomato+ (LysM+), Ly6G-, CD3-, CD19- cells (monocytes/macrophages) were sorted directly into QIAgen RLT+ buffer and used for SMART-seq2 cDNA library generation Full-length cDNAs and sequencing libraries were generated following the SMART-Seq2 protocol, as previously described (Picelli et al, 2014). Quality control (Agilent Technologies) and "tagmentation" - PCR-mediated adaptor addition and amplification of the adapted libraries was performed following the Nextera library preparation protocol (llumina Tagment DNA Enzyme and Buffer, Illumina #20034211; KAPA HiFi HotStart ReadyMix, Roche #07958935001; Nextera XT Index Kit v2 Set A, Illumina #15052163; Nextera XT Index Kit v2 Set D, Illumina #15052166), essentially as previously described (Baym et al, 2015). Libraries were sequenced (NextSeq2000 sequencing; Illumina) using 100 SE P2 and sequence information was extracted in FastQ format, using Illumina DRAGEN FASTQ Generation v3.8.4, producing on average approximately 38x10^6 (liver), 32x10^6 (WAT) and 43x10^6 (heart) reads per sample.