GSM1575621: 4C-seq: ind22_Kc_1; Drosophila melanogaster; OTHER
4C-seq in S2 and Kc cells was carried out as described by Splinter et al.63 with minor modifications as following: 50-100 million S2 cells or Kc cells, fixed, were used for two biological replicates. DpnII (NEB) was used as the primary and Csp6I (ThermoScientific) as the secondary restriction enzyme. For each viewpoint, two 50ul PCR reactions (8 cycles with 5C annealing temperature, followed by 18 cycles with 63C) with 160ng of template each were prepared and the combined products cleaned up by a one-step purification using 1.6 volumes of AMPure XP magnetic beads (Beckmann Coulter) according to manufacturer's instructions. These viewpoint libraries were eluted in 20ul elution buffer (10mM Tris-HCl pH 8.5) and the molarities for each individual sample were estimated using the bioanalyzer DNA 7500 kit (Agilent Technologies). All different viewpoint libraries for one biological replicate were mixed equimolarly and sequenced on separate lanes on an Illumina HiSeq2500 DNA sequencer.
GEO
GSM1575621