GSM5006286: AD2_9_old; Drosophila melanogaster; ATAC-seq
Midguts of ~100-140 flies were dissected and dissociated using 0.5% Trypsin-EDTA and 100,000 YFP-labeled intestinal stem cells were isolated using FACS. Cells were collected into PBS and immediately spun down at 4°C, washed and lysed to release nuclei, which were then transposed using the Nextera DNA Sample prep kit (Illumina) according to manufacturer's instructions. Transposed DNA was purified using MiniElute Reaction Cleaup kit (Qiagen). Transposed DNA was amplified using NEBNext High-Fidelity 2X PCR Master Mix (New England Biolabs). Libraries were quantified by qPCR to determine the cycle number at which each sample produced 25% of maximum fluorescent intensity. The fluorescent intensity was used to estimate the number of additional PCR cycles required for each sample.