GSM2633507: S2R+ rep 1; Drosophila melanogaster; Hi-C
Hi-C and library preparation was performed using the in-situ method as described previously (Rao et al., 2014) with minor modifications. Briefly, 1x10^6 cells were digested overnight at 37°C using 500 U of DpnII. After biotin filling, proximity ligation was carried out for 4 hr at 18°C with 2000 U T4 DNA Ligase, and after reverse-crosslinking, DNA was purified with ethanol precipitation and sheared to 300-400 bp fragments using a Covaris S220 sonicator. Ligation fragments with biotin were pulled-down using MyOne Streptavidin T1 beads (ThermoFisher Cat.N: 65602), end-repaired and adenylated as described. Fragments were then ligated with NEXTflex adaptors (Bioo Scientific, Cat.N: 514101) and amplified by PCR using KAPA HiFi Library Ampification Kit (Kapa Biosystems, Cat.N: KK2620) for 8 cycles. DNA size-selection (300-800 bp) was done using AMPure XP beads (Agencourt, Cat.N: A63881).
GEO Accession
GSM2633507