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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: 225, Reviews: 0 Write review Cached -
Promoter analysis by saturation mutagenesis
(Biological Procedures Online )
a powerful mutagenesis strategy that can be employed to define cis-acting elements at a molecular level. Technical details including primer design, saturation mutagenesis, construction of promoter libraries, phenotypic analysis, data analysis, and interpretation are discussed.
http://www.biologicalprocedures.com/bpo/arts/1/24/...
Added: Wed Sep 25 2002, Hits: 563, Reviews: 0 Write review Cached