GSM6448387: α-GFP, NPR1:GFP, +INA, MNase, rep3; Arabidopsis thaliana; ChIP-Seq
Seedlings were fixed with formaldehyde and then qeunched with glycine. After isolating nuclei from crosslinked sample, chromatin was fragmentated by sonication or micrococcal nuclease (MNase) treatment. NPR1:GFP or HAC1:mCherry proteins within the chromatin were immunoprecipitated by using anti-GFP or anti-RFP antibody, respectively. Genomic DNAs co-immunoprecipitated with NPR1:GFP or HAC1:mCherry were purified and used for sequencing. ChIP-seq libraries were prepared by using NEBNext® Ultra™ II DNA Library Prep with Sample Purification Beads (NEB, E7103) and NEBNext® Multiplex Oligos for Illumina® (NEB, E7335). 10 ng of DNA per sample was used for library-construction reaction. Prepared libraries were amplified by 9 cycles of PCR. For DNA samples collected from ChIP involving MNase digestion, sonication was additionally performed at mild condition before library preparartion.