GSM3601833: input_H3K9me2_ChIP-Seq_S.bicolor_60d; Sorghum bicolor; ChIP-Seq
Sixty-day fresh above-ground tissues were collected at the tillering stage for O. sativa vg. japonica, O. glaberrima, O. punctata, O. brachyantha, L. perrieri, and S. bicolor. Two-week fresh above-ground tissues at the seedling stage were also collected for O. sativa and O. brachyantha. ChIP experiments were performed as described in Gendrel et al. 2005 with minor modification. Briefly, 4g (for seedling stage) or 8g (for tillering stage) tissue were fine grounded in liquid nitrogen, followed by 3% formaldehyde fixation step. Nucleus were extracted and chromatin was sonicated into 300-500bp fragments in Bioruptor (Diagenode). Since each antibody has its own concentration and binding efficiency, we tested the optimal antibody volume for each antibodies. Typically we use 20ul antibody (1ug/ul) in a 350ul-400ul hybrid solution. The final DNA precipitation was used for library construction with the NEB Next N6240 kit (New England BioLabs) and decoded by Hiseq 2000 (Ilumina) with pair end 90bp strategy.
GEO Accession
GSM3601833