GSM8773804: brown adipose tissue, Male PGC-1a knockout, room temperature(25C); Mus musculus; ATAC-seq
Flash-frozen tissues were sent to Active Motif to perform the ATAC-seq assay. The tissue was manually dissociated, and nuclei were isolated by mechanical disruption. Isolated nuclei were quantified using a hemocytometer. A total of 100,000 nuclei were used for downstream processing. Nuclei were tagmented as previously described (Buenrostro et al., 2013), with modifications based on Corces et al. (2017), using the enzyme and buffer provided in the ATAC-Seq Kit (Active Motif). Tagmented DNA was purified using the MinElute PCR purification kit (Qiagen), followed by amplification with 10 cycles of PCR. The amplified DNA was further purified using Agencourt AMPure SPRI beads (Beckman Coulter). The resulting libraries were sequenced using paired-end 42 bp reads (PE42) on the NovaSeq 6000 sequencer (Illumina).