GSM2090079: pold2_H3K9me2; Arabidopsis thaliana; ChIP-Seq
Genomic DNAs were extracted from seedlings using the DNeasy Plant Mini Kit (QIAGEN)；total RNAs extracted from seedlings using the RNeasy Plant Mini Kit (QIAGEN) For ChIP-seq library, a 10-ng quantity of ChIP or input DNA was used to prepare a high-throughput sequencing library. End repair, dA-tailing, adapter ligation, and amplification were carried out using the NEBNext® ChIP-Seq Library Prep Master Mix Set for Illumina (E6240, NEB) according to the manufacturer’s protocol. Index primers used for each sample were as follows: ATCACG for WT-H3K27me3, CGATGT for WT-H3K4me3, TTAGGC for WT-H3K9me2, TGACCA for WT-H3, ACAGTG for WT-Input, GCCAAT for pold2-H3K27me3, CAGATC for pold2-H3K4me3, ACTTGA for pold2-H3K9me2, GATCAG for pold2-H3, and TAGCTT for pold2-Input. For Bisulfite-seq, a 2-μg quantity of genomic DNAs was sonicated (20 cycles of 30 s on, 30 s off, at low intensity) into 200-bp fragments with a Bioruptor (Diagenode, USA), end repaired, 3’-dA-tailed and ligated to methylated adapters. After purification, the ligated DNA fragments were treated with the EZ Methylation-Gold Kit (Zymo Research). The eluted bisulfite-treated DNAs were amplified using barcoded primers for sequencing (CGCTGT for the wild type and ACAGTG for the pold2 mutant)
GEO Accession
GSM2090079