GSM9140549: 35Spro:GFP input biological replicate 2; Arabidopsis thaliana; ChIP-Seq
Eight-day-old seedlings horizontally grown on 0.5 × MS medium plus 1% sucrose were used for crosslinking via 1% formaldehyde and the crosslinking was quenched by 125 mM glycine solution. Nuclei samples were extracted using HONDA buffer (20 mM HEPES KOH pH 7.4, 10 mM MgCl2, 440 mM sucrose, 1.25% Ficoll, 2.5% Dextran T40, 0.5% Triton X-100, 5 mM DTT, 5 mM phenylmethylsulfonyl fluoride [PMSF] and 1% Protease Inhibitor Cocktail [Sigma-Aldrich]). DNAs associated with chromatin were prepared following the protocol for Chromotek GFP-trap agarose beads (https://www.ptglab.com/products/pictures/pdf/ChIP_Apps_ Note_GFP-Trap_for_A_thaliana.pdf?redirect=chromotek) with some modifications. The Chip-seq libraries were constructed using NEBNext Ultra II DNA Library Prep Kit for Illumina following the manufacturer's instructions with minor modifications. Size selection was performed after PCR enrichment of adaptor-ligated DNA using SPRIselect beads (Beckman Coulter).