GSM8244458: Mutant fourth leaves,1-month-old,H3K27me3,rep2; Solanum lycopersicum; ChIP-Seq
ChIP-seq assays were performed according to Ramirez-Prado et al. (Ramirez-Prado et al, 2021) with extra modifications to the chromatin extraction step cross-linked plants were ground using gentleMACS (Miltenyi Biotec) with the 4C program in an extraction buffer (50mM HEPES-KOH pH 7.5, 150mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% sodium deoxycholate, 1% SDS) containing protease inhibitors. The mixture was filtered through a 50µm filter and left on ice for 10 minutes. After centrifuge for 10 minutes at 1500g at 4°C, the supernatant was removed and the pellet was resuspended in sonication buffer (50mM HEPES KOH pH 7.5, 150mM NaCl, 1mM EDTA, 1% Triton X-100, 0.2% SDS) containing protease inhibitors. 4ug of anti-H3K9me2 (Abcam, ab1220), anti-H3K27me1 (Millipore, 07–448), and anti-H3K27me3 (Millipore, 07-449) antibodies were used. ChIP-seq libraries were prepared from 10 ng of DNA using NEBNext Ultra II DNA Library Prep Kit for Illumina (NEB) according to the manufacturer's instructions.