Chromatin Immunoprecipitation Using Imbibed Seeds of Arabidopsis thaliana.
Methods Mol Biol
; 2830: 81-91, 2024.
Article
em En
| MEDLINE
| ID: mdl-38977570
ABSTRACT
Chromatin immunoprecipitation (ChIP) is used to analyze the targeting of a protein to a specific region of chromatin in vivo. Here, we present an instructive ChIP protocol for Arabidopsis imbibed seeds. The protocol covers all steps, from the sampling of imbibed seeds to the reverse crosslinking of immunoprecipitated protein-DNA complexes, and includes experimental tips and notes. The targeting of the protein to DNA is determined by quantitative PCR (qPCR) using reverse crosslinked DNA. The protocol can be further scaled up for ChIP-sequencing (ChIP-seq) analysis. As an example of the protocol, we include a ChIP-quantitative PCR (ChIP-qPCR) analysis demonstrating the targeting of PIF1 to the ABI5 promoter.
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Base de dados:
MEDLINE
Assunto principal:
Sementes
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Arabidopsis
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Proteínas de Arabidopsis
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Imunoprecipitação da Cromatina
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article