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Methods Mol Biol ; 2624: 55-72, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36723809

RESUMO

The chromatin immunoprecipitation coupled with the next-generation sequencing (ChIP-seq) is a powerful technique that enables to characterize the genomic distribution of chromatin-associated proteins, histone posttranslational modifications, and histone variants. However, in the absence of a reference control for monitoring experimental and biological variations, the standard ChIP-seq scheme is unable to accurately assess changes in the abundance of chromatin targets across different experimental samples. To overcome this limitation, the combination of external spike-in material with the experimental chromatin is offered as an effective solution for quantitative comparison of ChIP-seq data across different conditions. Here, we detail (i) the experimental protocol for preparing quality control spike-in chromatin from Drosophila melanogaster cells and (ii) the computational protocol to compare ChIP-seq samples with spike-in based on the use of the spikChIP software.


Assuntos
Sequenciamento de Cromatina por Imunoprecipitação , Histonas , Animais , Histonas/genética , Histonas/metabolismo , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Cromatina/genética , Imunoprecipitação da Cromatina/métodos , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Análise de Sequência de DNA/métodos
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