Endosperm-specific chromatin profiling by fluorescence-activated nuclei sorting and ChIP-on-chip.
Methods Mol Biol
; 1112: 105-15, 2014.
Article
em En
| MEDLINE
| ID: mdl-24478010
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.
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Assunto principal:
Cromatina
/
Núcleo Celular
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Análise Serial de Tecidos
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Imunoprecipitação da Cromatina
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Endosperma
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Citometria de Fluxo
Idioma:
En
Ano de publicação:
2014
Tipo de documento:
Article