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Protocols
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DNA methyltransferase-based single-molecule (MAPit) assay for mapping protein-DNA interactions
(Santhi Pondugula, Vamsi K. Gangaraju, Blaine Bartholomew and Michael P.Kladde)
This protocol describes a DNA methyltransferase-based single-molecule (MAPit) assay for mapping protein-DNA interactions in vitro'. MapIt can be used to footprint a variety of protein-DNA interactions, including nucleosomes, transcription factors and engaged polymerases. Here, they describe the application of this method to map nucleosome positions in biochemically reconstituted chromatin preparations either directly or after remodeling with the prototypical yeast ISW2 complex purified from budding yeast. This method should provide a powerful new approach for exploring mechanistic aspects of chromatin remodelling.
http://www.epigenome-noe.net/researchtools/protoco...
Added: Thu Feb 25 2010, Hits: 253, Reviews: 0 Write review Cached -
Peptide Pull-Down (PPD) Assay for Identification and Characterization of Histone PTM Effectors
(Andrew K. Roosand Joanna Wysocka)
To comprehend the complexity of epigenetic regulation, it is essential to not only catalogue histone PTMs and their patterns, but also to understand roles that histone PTMs and their effectors play in biological processes. An important component of this understanding will come through identification of histone PTM binding proteins. To this end, the peptide pull-down (PPD) assay provides a simple and effective tool to identify and characterize such reader proteins
http://www.epigenome-noe.net/researchtools/protoco...
Added: Thu Feb 25 2010, Hits: 264, Reviews: 0 Write review Cached -
Reverse Transfection for Gene Function Analysis
(Sabtini Lab, MIT)
This guide describes a microarray-based system for the functional analysis in mammalian cells of many genes in parallel. Mammalian cells are cultured on a glass slide printed in defined locations with solutions containing different DNAs. Cells growing on the printed areas take up the DNA, creating spots of localized transfection within a lawn of non-transfected cells. This method is called 'reverse transfection'.
http://web.wi.mit.edu/sabatini/pub/Downloadable_fi...
Added: Sat May 24 2003, Hits: 1862, Reviews: 0 Write review
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