title: GSM2667888: at-endosperm-input_replicate2_col_wt_x_col_4n-ahl10; Arabidopsis thaliana; ChIP-Seq
library_construction_protocol: INTACT-purified endosperm nuclei from 600-800 mg siliques were cross-linked 8 min with 1% formaldehyde and sonicated in 100 μl of NLB (Nuclei Lysis Buffer, 50 mM Tris-HCl pH8.0, 10 mM EDTA, 1% SDS, supplemented with Complete protease inhibitors) using a Bioruptor (Diagenode). 900 μl of ChIP dilution buffer (1.1% Triton X-100, 1.2 mM EDTA, 16.7 mM Tris-HCl pH 8.0, 167 mM NaCl, containing Complete protease inhibitors (Roche)) was added and chromatin was divided in different aliquots for an overnight incubation with antibodies. The following antibodies were used in this study: anti-Histone H3 (Sigma, #H9289,), H3K9me2 (Diagenode, #pAb‐060‐050). Antibodies IP were performed using Dynabeads (Invitrogen). DNA elution, de-crosslinking and purification steps were done using the IPure kit (Diagenode). ChIP‐seq libraries were generated using the Microplex library preparation kit (Diagenode) following the manufacturer's protocol with 1 ng of starting material.
attributes.tag: GEO Accession
attributes.value: GSM2667888