GSM5222662: ChIPseq of H3K4me3 in sunflower 13 days after anthesis developing seeds; Helianthus annuus; ChIP-Seq
The seeds were cross-linked with 1% formaldehyde by vacuum infiltration and quenched with 0.125 M glycine. After grinding, nuclei were isolated in extraction buffer (1% Triton X-100, 0.25 M sucrose, 10 mM MgCl2, 10 mM Tris-HCl pH 8.0 and protease inhibitors). Nuclei were pelleted by centrifugation and resuspended in lysis buffer (1% SDS, 50 mM Tris-HCl pH 8.0 and 10 mM EDTA). Then, nuclei were disrupted by sonication in Bioruptor obtaining genomic fragments from 200 to 600 bp. The rest of ChIP-Seq experiment was carried out as described previously by Villar and Köhler (2010) with minor modifications. 1ug of DNA/protein complexes was inmunoprecipitated with anti-H3K4me3 antibody. After crossliked reversal and DNA purification, the ChIP DNA was quantified with the Qubit HS dsDNA kit (Invitrogen). Illumina libraries for sequencing were generated, and the libraries were sequenced using the HiSEq 2000 Sequencing System (Illumina), following the manufacturer's instructions.
GEO
GSM5222662