alias: E-MTAB-1043:EAS488_6
title: ChIP-seq of LHP1 and H3K27me3 distribution in Arabidopsis thaliana
design: ChIP-seq of LHP1 and H3K27me3 distribution in Arabidopsis thaliana
library_construction_protocol: A. thaliana plants of the accessions Col and Ler were used as genetic background. For intraspecific crossing, Col and Ler plants were grown on soil in LD conditions. Five flower buds on the primary shoot and two side shoots were emasculated and manually cross pollinated, while all other flower buds were removed. The seeds of each genotype were pooled and the success rate of the crosses was determined by PCR using primers that detected an insertion between Ler and Col (fw: 5-ctggagatcatccaacaaagg-3, rv: 5-ggcaatggaatgggctggtc-3). Seed pools with less than 10 % maternal contamination were used in the F1 hybrid studies. Hybrids of the F1 generation were grown in LD (16-hours light, 8-hours dark) at 20C constant temperature on Murashige and Skoog (GM) medium supplemented with 1 % sucrose after stratification at 4C for 2-4 days to synchronize germination. Light was provided by fluorescent tubes complemented by incandescent bulbs to increase the proportion of far red light. Nuclear extraction and ChIP experiments were performed as described in J.J. Reimer and F. Turck; Genome-wide mapping of protein-DNA interaction by chromatin immunoprecipitation and DNA microarray hybridization (ChIP-chip). Part A: ChIP-chip molecular methods, Methods Mol Biol, 631, 139-160, 2010.
identifiers.namespace: University of Cologne, Cologne Center for Genomics
identifiers.id: E-MTAB-1043:EAS488_6