GSM9518339: snRNAseq, Epigonadal Adipocytes; Mus musculus; RNA-Seq
Thymic capsules were cleaned of directly attached perithymic fat and connective tissues, rinsed a final time with PBS, and transferred to a 15mL conical tube for enzymatic digestion with DNase I and Collagenase II in DMEM-F12 10% FBS. Samples were fractionated by sequential centrigugation steps, and the topmost white layer containing mature, floating adipocytes was obtained. Nuclei were isolated as previously described (10X Genomics, Demonstrated Protocol CG000366 Rev D) followed by library preparation and joint RNAseq + ATACseq (6mo F) or RNAseq alone (4mo M) using the 10x Genomics platform by the University of Utah High Throughput Genomics Core. 5,000-10,000 nuclei were targeted for snRNAseq. Libraries were prepared using Library Construction Kit B reagents and sequenced on an Illumina NovaSeq X instrument targeting at least 100 million paired end reads (GEX) and at least 300 million paired end reads (ATAC) of 150 bp.