GSM3466753: SOB3-GFP ChIP replicate 2; Arabidopsis thaliana; ChIP-Seq
Frozen samples were ground using mortar and pestle. Ground tissue was mixed with fixation buffer and formaldehyde added to 1% final concentration. Fixation was stopped after 10 minutes by addition of glycine. Solution was filtered twice through miracloth and then centrifuged to pellet cells.  Pellet was resuspended in extraction buffer containing 1% Triton X-100 to lyse cells and centrifuged to pellet nuclei. Pellet with nuclei was resuspended in nuclei separation buffer containing 1.7 M sucrose and centrifuged to separate nuclei from debris. Nuclei were resuspended in nuclei lysis buffer containing 1% SDS and flash frozen twice in liquid nitrogen in order to lyse nuclei.  Subsequently, chromatin was sonicated in a Bioruptor (Diagenode) to an average size of approximately 200-400 bp. Libraries were constructed from input or ChIPed DNA using the TruSeq ChIP Library Preparation Kit (Illumina) according to manufacturer's instructions, with the exception of the "Purify Ligation Products" step, which was skipped. Library quality was validated using the Agilant 2200 TapeStation with D1000 ScreenTape & Reagents. Qubit dsDNA HS Assay Kit was used with the Qubit 2.0 Fluorometer (Invitrogen) for quantitation of DNA.
GEO Accession
GSM3466753