Chip-Seq Arabidopsis thaliana Spot-ARM_2 replicate 2
7-day-old seedlings were washed with sterile water and cross-linked with 2% formaldehyde for 10 minutes under vacuum, then cross-linking was quenched with 2 M glycine (final concentration 0.125 mM). Chromatin was isolated from cross-linked seedlings following Saleh et al. (2008) up to nuclei pellet preparation. Nuclei pellets were incubated on ice for 20 min with nuclei lysis buffer and sonicated for chromatin fragmentation. Chromatin was immunoprecipitated using Spot-Trap Magnetic Agarose (ChromoTek) and samples were eluted according manufacturer's recommended protocol DNA from input. After reverse crosslinking and RNaseA and proteinase K treatment, DNA was purifed using AMPure XP magnetic beads. Immunoprecipitated samples (>5 ng per sample) were checked using the 5200 Fragment Analyzer (Agilent). Libraries were prepared with the Watchmaker DNA library prep kit and UMI-containing adapters, and passed the QC with sufficient concentration >10ng/ul, fragment size and absence of adapter dimers. Libraries were sequenced on the MGI G400 instrument with a DNBSEQ G400RS PE150 high output sequencing set. Input samples were sequenced to a depth of 40 million reads per replicate, while pulldown samples were sequenced to 10 million reads per replicate.