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Protocol for using heterologous spike-ins to normalize for technical variation in chromatin immunoprecipitation.
STAR Protoc. 2:100609 (2021)
Quantifying differential genome occupancy by chromatin immunoprecipitation (ChIP) remains challenging due to variation in chromatin fragmentation, immunoprecipitation efficiencies, and intertube variability. In this protocol, we add heterologous spike-ins from Drosophila chromatin as an internal control to the mice chromatin before immunoprecipitation to normalize for technical variation in ChIP-qPCR or ChIP-seq. The choice of spike-in depends on the evolutionary conservation of the protein of interest and the antibody used. For complete details on the use and execution of this protocol, please refer to Greulich et al. (2021).
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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
Cell Biology ; Chipseq ; Chromatin Immunoprecipitation (chip) ; Molecular Biology ; Sequence Analysis
e-ISSN
2666-1667
Journal
STAR Protocols
Quellenangaben
Volume: 2,
Issue: 3,
Article Number: 100609
Non-patent literature
Publications
Reviewing status
Peer reviewed
Institute(s)
Institute of Diabetes and Cancer (IDC)