title: GSM5915105: NLP2-T30_rep2; Arabidopsis thaliana; ChIP-Seq
library_construction_protocol: Chromatin extraction and immunoprecipitation was performed essentially as described in (Bouyer et al., 2018; https://doi.org/10.1371/journal.pgen.1007797) with minor modifications. Briefly, 800 mg of liquid nitrogen-ground seedlings were subject to double crosslinking (2.5mM Di(N-succinimidyl) glutarate 60 min followed by 1% formaldehyde 7 min). Chromatin fragmentation was done using a Covaris S220 ultra-sonicator with the following parameters: 12 min with 5% duty cycle, 105W peak power and 200 cycles per burst. Chromatin immunoprecipitation was done using 3 ug of anti-GFP antibodies (ab290,Abcam) pre-bound to proteinA/G-coupled magnetic beads. For each replicate, 250 pg of immunoprecipitated (IP) and genomic (INPUT) DNA were used to prepare libraries with the MicroPlex Library Preparation kit (Diagenode). Quality of libraries was validated using Agilent / Advanced Analytical fragment analyzer. Multiplexed libraries were sequenced using a NextSeq 500  system (Illumina) with paired-ends 41 bp reads.