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Book chapter2014

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

Weinhofer, Isabelle; Köhler, Claudia

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
ISBN: 978-1-62703-773-0 (electr.)
Publisher: Springer

    UKÄ Subject classification

    Plant Biotechnology
    Biochemistry and Molecular Biology

    Publication identifier

    DOI: https://doi.org/10.1007/978-1-62703-773-0_7

    Permanent link to this page (URI)

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