GSM2586373: Fruit H3K27me3 replicate 2; Cucumis melo var. cantalupo; ChIP-Seq
Total RNA were extracted from leaves, root, fruit, male and female flowers using the Nucleospin RNA kit (Macherey-Nagel). ChIP assays were performed using anti-H3K9ac (Millipore) or anti-H3K27me3 (Millipore) antibodies. Chromatin was reverse-crosslinked by adding 20μL of NaCl 5M and incubated over-night at 65°C. Reverse-cross-linked DNA was submitted to RNase and proteinase K digestion, and extracted with phenol-chloroform. DNA was ethanol precipitated in the presence of 20μg of glycogen and resuspended in 50μL of nuclease-free water (Ambion) in a DNA low-bind tube. 10 ng of IP or input DNA was used 501 for ChIP-Seq library construction using NEB-Next Ultra II DNA Library Prep Kit for Illumina (New England Biolabs) according to manufacturer’s recommendations. For all libraries, twelve cycles of PCR were  used. The quality of the libraries was assessed with Agilent 2100 Bioanalyzer (Agilent), and  the libraries were subjected to high-throughput sequencing by Illumina Sequencing technology.
GEO Accession
GSM2586373