GSM4160824: NPP1_12h_Rep1; Arabidopsis thaliana; RNA-Seq
RNA was extracted from samples 12 hours after induction from three independent biological replicates using TRIzol reagent (Invitrogen), treated with Turbo DNAse (Thermo Fisher Scientific). For chromatin immunoprecipitation, leaf material at 12 hours after induction was collected in PBS (137 mM NaCl, 2.7 mM KCl, 4.3 mM Na2HPO4, 1.4 mM KH2PO4) supplemented with 0.1% Tween and cross‐linked using 1% formaldehyde for 10 min under vacuum. Cross-linking was stopped by glycine at final concentration 125 mM for 5 min under vacuum. Leaves were rinsed with water, blotted dry and frozen in liquid nitrogen. About 200 mg of liquid nitrogen-frozen leaves were used for crude nuclei extraction according to (Mozgová et al., 2015). ChIP experiments were performed were done as previously described (Mozgová et al., 2015) from three independent biological replicates. RNA sequencing libraries were prepared using TruSeq RNA Library Preparation Kit v2 (Illumina) and 18 samples were pooled for sequencing in one lane of the Illumina HiSeq2000 platform. ChIP-seq libraries were prepared with Microplex Library Preparation kit v2 (Diagenode) with 2 ng of eluted DNA as starting material and 12 samples were pooled for sequencing in one lane of Ilumina HiSeq 2500 platform with Single reads of 50 bp length.
GEO Accession
GSM4160824