Cargando…

Microplate-based chromatin immunoprecipitation method, Matrix ChIP: a platform to study signaling of complex genomic events

The chromatin immunoprecipitation (ChIP) assay is a major tool in the study of genomic processes in vivo. This and other methods are revealing that control of gene expression, cell division and DNA repair involves multiple proteins and great number of their modifications. ChIP assay is traditionally...

Descripción completa

Detalles Bibliográficos
Autores principales: Flanagin, Steve, Nelson, Joel D., Castner, David G., Denisenko, Oleg, Bomsztyk, Karol
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2241906/
https://www.ncbi.nlm.nih.gov/pubmed/18203739
http://dx.doi.org/10.1093/nar/gkn001
_version_ 1782150554208698368
author Flanagin, Steve
Nelson, Joel D.
Castner, David G.
Denisenko, Oleg
Bomsztyk, Karol
author_facet Flanagin, Steve
Nelson, Joel D.
Castner, David G.
Denisenko, Oleg
Bomsztyk, Karol
author_sort Flanagin, Steve
collection PubMed
description The chromatin immunoprecipitation (ChIP) assay is a major tool in the study of genomic processes in vivo. This and other methods are revealing that control of gene expression, cell division and DNA repair involves multiple proteins and great number of their modifications. ChIP assay is traditionally done in test tubes limiting the ability to study signaling of the complex genomic events. To increase the throughput and to simplify the assay we have developed a microplate-based ChIP (Matrix ChIP) method, where all steps from immunoprecipitation to DNA purification are done in microplate wells without sample transfers. This platform has several important advantages over the tube-based assay including very simple sample handling, high throughput, improved sensitivity and reproducibility, and potential for automation. 96 ChIP measurements including PCR can be done by one researcher in one day. We illustrate the power of Matrix ChIP by parallel profiling 80 different chromatin and transcription time-course events along an inducible gene including transient recruitment of kinases.
format Text
id pubmed-2241906
institution National Center for Biotechnology Information
language English
publishDate 2008
publisher Oxford University Press
record_format MEDLINE/PubMed
spelling pubmed-22419062008-02-21 Microplate-based chromatin immunoprecipitation method, Matrix ChIP: a platform to study signaling of complex genomic events Flanagin, Steve Nelson, Joel D. Castner, David G. Denisenko, Oleg Bomsztyk, Karol Nucleic Acids Res Methods Online The chromatin immunoprecipitation (ChIP) assay is a major tool in the study of genomic processes in vivo. This and other methods are revealing that control of gene expression, cell division and DNA repair involves multiple proteins and great number of their modifications. ChIP assay is traditionally done in test tubes limiting the ability to study signaling of the complex genomic events. To increase the throughput and to simplify the assay we have developed a microplate-based ChIP (Matrix ChIP) method, where all steps from immunoprecipitation to DNA purification are done in microplate wells without sample transfers. This platform has several important advantages over the tube-based assay including very simple sample handling, high throughput, improved sensitivity and reproducibility, and potential for automation. 96 ChIP measurements including PCR can be done by one researcher in one day. We illustrate the power of Matrix ChIP by parallel profiling 80 different chromatin and transcription time-course events along an inducible gene including transient recruitment of kinases. Oxford University Press 2008-02 2008-01-18 /pmc/articles/PMC2241906/ /pubmed/18203739 http://dx.doi.org/10.1093/nar/gkn001 Text en © 2008 The Author(s) http://creativecommons.org/licenses/by-nc/2.0/uk/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods Online
Flanagin, Steve
Nelson, Joel D.
Castner, David G.
Denisenko, Oleg
Bomsztyk, Karol
Microplate-based chromatin immunoprecipitation method, Matrix ChIP: a platform to study signaling of complex genomic events
title Microplate-based chromatin immunoprecipitation method, Matrix ChIP: a platform to study signaling of complex genomic events
title_full Microplate-based chromatin immunoprecipitation method, Matrix ChIP: a platform to study signaling of complex genomic events
title_fullStr Microplate-based chromatin immunoprecipitation method, Matrix ChIP: a platform to study signaling of complex genomic events
title_full_unstemmed Microplate-based chromatin immunoprecipitation method, Matrix ChIP: a platform to study signaling of complex genomic events
title_short Microplate-based chromatin immunoprecipitation method, Matrix ChIP: a platform to study signaling of complex genomic events
title_sort microplate-based chromatin immunoprecipitation method, matrix chip: a platform to study signaling of complex genomic events
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2241906/
https://www.ncbi.nlm.nih.gov/pubmed/18203739
http://dx.doi.org/10.1093/nar/gkn001
work_keys_str_mv AT flanaginsteve microplatebasedchromatinimmunoprecipitationmethodmatrixchipaplatformtostudysignalingofcomplexgenomicevents
AT nelsonjoeld microplatebasedchromatinimmunoprecipitationmethodmatrixchipaplatformtostudysignalingofcomplexgenomicevents
AT castnerdavidg microplatebasedchromatinimmunoprecipitationmethodmatrixchipaplatformtostudysignalingofcomplexgenomicevents
AT denisenkooleg microplatebasedchromatinimmunoprecipitationmethodmatrixchipaplatformtostudysignalingofcomplexgenomicevents
AT bomsztykkarol microplatebasedchromatinimmunoprecipitationmethodmatrixchipaplatformtostudysignalingofcomplexgenomicevents