title: GSM4222136: pupal wing, D2_66hAPF_rep3; Drosophila melanogaster; ATAC-seq
library_construction_protocol: The protocol is modified from Omni-ATACseq. 24 pupal wings were then dissected, rinsed in cold PBS for twice, and moved into 100µl cold 1x lysis buffer. The wings were further cut into 3-4 pieces and transferred into a 2ml Dounce homogenizer. They were then disrupted by 12 strokes using pestle A, rest on ice for 5min, then 20 strokes using pestle B. After additional 10min incubation on ice, 900µl 1x wash buffer was added. 20ml syringe and 20 ½ gauge needle were employed here to separate cells from the wing cuticle. The mixture was then filtered with 40µM strainer , and centrifuged at 4°C, 1000g for 10min. Pelleted nuclei were gently resuspended with 45µl ultrapure water and counted using hemocytometer. 50000 nuclei were then centrifuged at 4°C, 1000g for 10min, resuspended with 8ml 2x Tagment DNA buffer. The tagmentation reaction followed the standard ATAC-seq protocol with minor modifications: 10ul 2X TD buffer with nuclei; 2ul Tagment DNA Enzyme; 8ul ultrapure water. Libary was prepaire using Illunmina standard protocol.
attributes.tag: GEO Accession
attributes.value: GSM4222136