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  • Tissue-Specific In Vivo Biotin Chromatin Immunoprecipitation with Sequencing in Zebrafish and Chicken.

Tissue-Specific In Vivo Biotin Chromatin Immunoprecipitation with Sequencing in Zebrafish and Chicken.

STAR protocols (2020-10-29)
Martyna Lukoseviciute, Irving T C Ling, Upeka Senanayake, Ivan Candido-Ferreira, Gunes Taylor, Ruth M Williams, Tatjana Sauka-Spengler
ABSTRACT

Chromatin immunoprecipitation with sequencing (ChIP-seq) has been instrumental in understanding transcription factor (TF) binding during gene regulation. ChIP-seq requires specific antibodies against desired TFs, which are not available for numerous species. Here, we describe a tissue-specific biotin ChIP-seq protocol for zebrafish and chicken embryos which utilizes AVI tagging of TFs, permitting their biotinylation by a co-expressed nuclear biotin ligase. Subsequently, biotinylated factors can be precipitated with streptavidin beads, enabling the user to construct TF genome-wide binding landscapes like conventional ChIP-seq methods. For complete details on the use and execution of this protocol, please see Lukoseviciute et al. (2018) and Ling and Sauka-Spengler (2019).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Glycine, BioUltra, Molecular Biology, ≥99.0% (NT)
Sigma-Aldrich
Lithium chloride, Molecular Biology, ≥99%
Sigma-Aldrich
HEPES, ≥99.5% (titration)
Sigma-Aldrich
Sodium deoxycholate, ≥97% (titration)
Sigma-Aldrich
Triton X-100, laboratory grade
Roche
Glycogen, from mussels