title: H3K9ac ChIP-seq of Foxtail millet (Setaria italica) genotype IC4/ IC403579 - Control
design: The Seeds of dehydration and salinity tolerant IC-403579 (IC4) and sensitive IC-480117 (IC41) cultivars of foxtail millet were sown in a plant growth chamber with the following conditions; 281C day/ 231C night/ 705% relative humidity with a photoperiod of 14 hr and a photosynthetic photon flux density of 500 mol m-2 s-1. For dehydration stress, twenty-one days old seedlings were treated with 20% polyethylene glycol 600 (PEG-6000). The dehydration stressed samples were collected at 24 hr after PEG treatments. The untreated samples were also collected as controls (0hr). For ChIP assay, 4g of the samples were crosslinked and crushed to fine powder using liquid nitrogen. This powder was resuspended in the nuclei isolation buffer, homogenize and filtered through four layers of cheesecloth. The filtered slurry was centrifuged at 11,000 g for 20 min at 4C. Then the nuclei pellet was resuspended in nuclei lysis buffer and subjected to sonication of chromatin after which these samples were centrifuged at 13,800 g for 10 min at 4C and supernatant was collected and used for the ChIP-assay. The collected supernatant was pre-cleared by mixing of 40 l Protein A agarose beads and incubated for 1 hr at 4C with gentle agitation. Then, the slurry was centrifuged at 16000 g for 2 minutes at 4C and the supernatant was collected. This supernatant was divided into ChIP, input control and no-antibody control (No-Ab, negative control). The ChIP-samples were immunoprecipitated by anti-H3K9Ac antibody using 60 l protein A agarose beads followed by washing with low and high salt, LiCl and TE buffer. Then, the ChIP-DNA was eluted by freshly prepared elution buffer (1% SDS and 0.1M NaHCO3) and followed by reverse cross-linking for overnight at 65C after adding 20 l of 5M NaCl. The proteinase K treatment was performed at 55C for 1hr followed by DNA purification. Then, ChIP and input DNA samples of both the cultivars were sequenced.