Endosperm-specific chromatin profiling by fluorescence-activated nuclei sorting and ChIP-on-chip

Abstract

Cell-type-specific analysis of gene expression and chromatin profiling requires the isolation of discrete cell populations from complex pools. However, until now this most critical step has been labor intensive and technical challenging. Here, we describe a rapid protocol based on fluorescence-activated cell sorting (FACS) for cell-type-specific RNA and chromatin profiling. We detail how to isolate nuclei from Arabidopsis inflorescence and silique homogenates and how to purify endosperm nuclei labeled by nuclear-targeted green fluorescent protein using FACS. The purified fluorescent endosperm nuclei can be further used for chromatin immunoprecipitation (ChIP) followed by hybridization to high-resolution whole-genome tiling microarrays (ChIP-on-chip) or transcriptional profiling.

Published in

Methods in Molecular Biology
2014, volume: 1112, number: 1112, pages: 105-115
Title: Plant Epigenetics and Epigenomics: Methods and Protocols
Publisher: Springer

SLU Authors

• Köhler, Claudia

  • Department of Plant Biology, Swedish University of Agricultural Sciences
    

UKÄ Subject classification

Plant Biotechnology
Biochemistry and Molecular Biology

Publication identifier

• DOI: https://doi.org/10.1007/978-1-62703-773-0_7
• ISBN: 978-1-62703-773-0 (electr.)

Permanent link to this page (URI)

https://res.slu.se/id/publ/66863