ATAC-Seq of Bicyclus anynana: early pupal (3 to 6 hr after pupation) forewing Cu1 eyespot centre
Small wing tissue containing eyespot centre in Cu1 sector was dissected from early pupa (between 3-6 hr after pupation).  50 tissues were pooled together (25 individuals) for each replicate. Samples were snap frozen immediately after dissection using liquid N2 and stored in -80C until further use. ~80,000 nuclei were used for each library. Nuclei isolation and library preparation were done with little modification as described in Omni-ATAC protocol (Corces et al, 2017). Sequencing was performed at AIT Novogene, Singapore; Reads from the sample were aligned to the Bicyclus anynana (BaGv2) genome using bowtie. Peak call was done using F-seq.