GSM3721303: silique.gfp.wt_rep2; Arabidopsis thaliana; ChIP-Seq
Crosslinking of plant material was done by collecting 600 mg of 2DAP PHE1::PHE1-GFP siliques, and vacuum infiltrating them with a 1% formaldehyde solution in PBS. The vacuum infiltration was done for two periods of 15 min, with a vacuum release between each period. The crosslinking was then stopped by adding 0.125 mM glycine in PBS and performing a vacuum infiltration for a total of 15 min, with a vacuum release each 5 min. The material was then ground in liquid nitrogen, resuspended in 5mL Honda buffer (Moreno-Romero et al., 2017), and incubated for 15 min with gentle rotation. This mixture was filtered twice through Miracloth and one time through a CellTrics filter (30 µm), after which a centrifugation for 5 min, at 4°C and 1,500 g was performed. The nuclei pellet was then resuspended in 100 µL of nuclei lysis buffer(Moreno-Romero et al., 2017), and the ChIP protocol was continued as described before(Moreno-Romero et al., 2017). GFP Tag Antibody (A-11120, Thermo Fisher Scientific) was used in this ChIP. Libraries were prepared using the Ovation Ultralow v2 Library System (NuGEN), with a starting material of 1 ng, following the manufacturer's instructions
GEO Accession
GSM3721303