GSM1463808: At_BLJ_YFP_1; Arabidopsis thaliana; ChIP-Seq
Roots were collected on nylon mesh and fixed for 15 min with 1.6% formaldehyde in PBS-EDTA. Roots were rinsed, dried, ground with mortar and pestle in liquid N, and dissolved in same volume of 50 mM Tris-Cl pH 7.5, 150 mM NaCl, 2.5 mM EDTA, 2mM PMSF, 2x Complete Protease Inhibitors, 0.5% Sarkosyl. This mixture was processed in Bioruptor for 15 min at 4 °C. Supernatant was diluted 6 times with 50 mM Tris-Cl pH 7.5, 150 mM NaCl, 2.5 mM EDTA, 2mM PMSF, 2x Complete Protease Inhibitors and immunoprecipitated with a-GFP AbCam290 at 4 °C o/n, Pierce Protein G beads and Pierce Spin Cups. DNA purification was performed by standard RNAse-A and proteinase K treatments followed by Qiagen MinElute kit Immunoprecipitated purified DNA was linearly amplified and processed with Nextera DNA Sample Prep kit.  Briefly, genomic regions were fragmented and adaptors ligated using Nextera transposon insertion system. These fragments were make into libraries of ~100 bp insert size. Adaptors cointain barcodes and thus samples weredistincly barcoded. Libraries were mixed and sequenced in one lane.  50 bp of single ends were sequenced through Illumina High Sequencing.