GSM2665528: HD2B:GFP flg; Arabidopsis thaliana; ChIP-Seq
[ChIP-seq] ChIP assays were performed on 14-day-old in vitro seedlings using anti-GFP , IgG control, or anti-H3K9ac antibodies. Briefly, after plant material fixation in 1% (v/v) formaldehyde, tissues were homogenized, nuclei isolated and lysed. Cross-linked chromatin was sonicated using a water bath Bioruptor UCD-200 (Diagenode, Liège, Belgium) (30 s on/30 s off pulses, at high intensity for 60 min). Protein/DNA complexes were immunoprecipitated with antibodies, overnight at 4°C with gentle shaking, and incubated for 1h at 4°C with 50 μL of Dynabeads Protein A (Invitrogen, Ref. 100-02D). Immunoprecipitated DNA was then recovered using the IPure kit (Diagenode, Liège, Belgium) and analyzed by quantitative real-time PCR. An aliquot of untreated sonicated chromatin was processed in parallel and used as the total input DNA control. [ChIP-seq] 10 ng of IP or input DNA was used 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. [RNA-seq] Total RNA were extracted from seedlings with the RNeasy MiniPrep kit (Qiagen), according to the manufacturer's instructions. [RNA-seq] Transcriptomic analyses were performed RNA-seq at the URGV plateform (Evry, France) with Illumina RNA-seq preparation kit. Three biological replicates were run for each condition.
GEO
GSM2665528
GEO Accession
GSM2665528