GSM9203568: ref6-5, 10DAG, H2Aub; Arabidopsis thaliana; ChIP-Seq
ChIP experiments were performed on 1 g of 10-day-old seedling tissue and 5 g of 48-hour imbibed seeds (48 HAI) fixed in 1% formaldehyde. Chromatin was extracted from the fixed tissue and fragmented using a Bioruptor Pico (Diagenode). The sheared chromatin was immunoprecipitated overnight using the following antibodies and dilutions: anti-H3K27me3 (1:300, Millipore 07-449), anti-HsH2Aub (1:100, CST 8240S), anti-H3K27me1 (1:150, Millipore 07–448) or anti-HTA9 (1:300) antibodies. Immunocomplexes were captured using Protein-A Sepharose beads CL-4B (GE Healthcare) and crosslinking was reversed overnight at 65ºC. Final ChIP DNA was purified with MicroChIP DiaPure columns kit (Diagenode C03040001). Libraries for ChIP-seq samples were prepared with TruSeq ChIP Library Preparation Kit (Illumina) and sequenced using Illumina NextSeq with single-end 75 bp reads