title: GSM7819302: DEK2,Input,ChIP; Arabidopsis thaliana; ChIP-Seq
library_construction_protocol: ChIP-seq assays were performed according to Ramirez-Prado et al. (Ramirez-Prado et al, 2021) with minor modifications. Plant materials were cross-linked with 1%(v/v) formaldehyde under vacuum. Chromatin was isolated and fragmented by sonication using Covaris S220 sonicator. Later, the sonicated protein/DNA was incubated with anti-GFP antibodies (Abcam, ab290) overnight at 4 °C with gentle shaking, and then incubated with 50 μL of Dynabeads Protein A (Invitrogen, Ref. 100-02D) for 1h at 4 °C. After beads washing, samples were reverse crosslinked, treated with RNase A+T1 and proteinase K. Immunoprecipitated DNA was then recovered by phenol:chloroform extraction and ethanol precipitation. DNA was quantified with Qubit High Sensitivity kit (Life technologies). ChIP-seq libraries were performed using 10ng of immunoprecipitated DNA and NebNext DNA Ultra II library preparation kit according to manufacturer's instructions.