GSM1686148: hpo-gfp_WT; Neurospora crassa; ChIP-Seq
Mycelia were cross-linked for 10 min in 0.5% formaldehyde for histone modification ChIP or 30 min in 1% formaldehyde for GFP ChIP.  Tissue was disrupted by sonication for thirty pulses before chromatin was sheared using a Bioruptor (Diagenode) for 15 min with a cycle of 30 sec on followed by 30 sec off, at high power.  DNA was purified using the QIAquick PCR purification kit  and eluted in 25 μl of water. Approximately 10 ng of DNA was used to generate ChIP-Seq libraries.  Each library was prepared using the Illumina Tru-Seq kits A and B (Illumina, IP-202-1012 and IP-202-1024) according to the manufacturer's instructions.  “Invisible” fragments between 250-400 bp were excised and purified using the MinElute gel extraction kit (Qiagen, 28606).  Final libraries were PCR-amplified using one cycle at 98 °C for 30 sec, 18 cycles at 98 °C for 10 sec, 60 °C for 30 sec and 72 °C for 30 sec and a final extension at 72 °C for 5 min.
GEO Accession
GSM1686148