title: GSM9326780: dss_ctrl2; Drosophila melanogaster; ATAC-seq
library_construction_protocol: Illumina Tagment DNA enzyme and buffer kit (REF 20034210 ATAC-seq libraries were manufactured according to the original protocol (Buenrostro JB, et.al 2015). Briefly, 50-75K GFP positive sorted cells were collected in ATAC lysis buffer (10mM Tris-HCl ph 7.4, 10mM NaCl, 3mM MgCl2, 0.1% Igepal CA-630). Cells were pelleted 10' x 500g, 4oC, supernatant discarded and subsequently gently suspended in 700ul ice-cold ATAC lysis buffer. After an additional 10' x 500g, 4oC spin, lysis buffer was removed with a 1000p pipette leaving 22.5ul covering the nuclei. We added 2.5ul Tagmentation enzyme and 25ul 2xTagmentation Buffer (Illumina Tagment DNA enzyme and buffer kit (REF 20034210). Fragmentation reactions were performed using a bioSan TS-100 Thermo-Shaker at 37oC for 40' shaking at 800RPM. Initial 5 cycle PCR was performed with Phusion High-Fidelity PCR Master Mix (NEB M0532S) using the Illumina Set C plate (REF 20026934) in total volume of 50ul. 5ul were used to estimate product cycle saturation with SYBR green in Biorad qPCR. Final libraries were produced after 13-14 total cycles of amplification and analyzed on a Bioanalyzer DNA chip.