Standard NextSeq 500 paired-end Sequencing; Whole genome sequencing of TAD B MiMIC line (Drosophila melanogaster)
Whole genome sequencing of TAD B MiMIC line (Drosophila melanogaster)
50 adult flies grown at 25°C were snap-frozen in liquid nitrogen. ; Flies were grinded on dry ice in an Eppendorf tube with a pestle. Grinded flies were resuspended in 250 µl of Buffer A (100 mM Tris-HCl pH7.5, 100 mM EDTA, 100 mM NaCl, 1% SDS) with 5 µl of proteinase K 20 µg/µl. The sample was incubated at 65°C for 3 hours and at 95°C for 10 min, then spun for 2 min at room temperature. The supernatant was transferred to a new tube and incubated at 37°C for 45 min with 2 µl of RNaseA 100 mg/ml. The DNA was extracted by two rounds of standard phenol-chloroform extraction, precipitated with ethanol, and resuspended in 150 µl of Tris-HCl 10 mM. ; 500 ng DNA were sonicated in 150 μl Tris-HCl 10 mM (5 pulses, 30'' on 30'' off) to obtain 200-400 bp fragments. DNA was size-selected with AMPure beads 1.4x. Library preparation was done with the NEBNext Ultra II DNA Library Prep kit according to manufacturer's instructions.