GSM8574684: Dmel, genitalia, 30 hAPF; Drosophila melanogaster; ATAC-seq
Pupae were dissected in ice-cold 1XPBS, followed by incubation in lysis buffer (10 mM Tris-HCL, pH = 7.5; 10 mM NaCl; 3 mM MgCl2; 0.1% IGEPAL and 0.1% digitonin). After the lysis was stopped, the nuclei were collected by centrifuging at 500g for 10 minutes. 50000 nuclei were resuspended in Tagmentation Mix [25 ul tagmentation buffer (20 mM Tris-CH2COO-, pH = 7.6; 10 mM MgCl2; 20 % Dimethylformamide); 2.5 Tn5 transposase; 22.5 ul H2O] and incubated for 30 minutes at 37 ºC. 3 ul of 2 M NaAC, pH = 5.2 was added to the sample and then purified using a QIAGEN MinElute Kit ATAC-seq library amplification was performed using NEBNext High Fidelity 2X PCR Mix with primers indexed with primers compatible with Nextera adaptors from Diagenode. 150 bp paired-end sequencing was carried out by Novogene (UK).