GSM8045950: ATACseq_sticklebackPrimarySpermatocytesGenomicControl_FreshwaterPool3; Gasterosteus aculeatus; ATAC-seq
For each ATAC-seq replicate (Buenrostro et al (2015) 109:2191-99), the testes of three males were pooled and dissociated with 0.07U/ml Liberase (Sigma). The isolation of 50000 primary spermatocytes per lysate was carried out by flow cytometry using a BD FACSMelody Cell Sorter (BD Biosciences) based on FSC-A and SSC-A (separating 4C primary spermatocytes, 2C secondary spermatocytes, 1C spermatids/sperm by size and complexity). Cell membranes were lysed with cold 10mM Tris pH8; 10mM NaCl; 3mM MgCl2; 0.1% NP-40 and nuclei were isolated by centrifugation. Tn5 was applied onto the nuclei in standard TAPS-DMF buffer for 30min +37'C with shaking (1000rpm), the tagged DNA was cleaned with the Min Elute kit (Qiagen) and processed to a sequencing library. A control library of naked genomic DNA was also made from each testes pool.  The libraries were paried-end sequenced on an Illumina platform.