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  Fluorescence detection of nucleic acids and proteins in multi-component crystals.

Kettenberger, H., & Cramer, P. (2006). Fluorescence detection of nucleic acids and proteins in multi-component crystals. Acta Crystallographica Section D: Biological Crystallography, 62(2), 146-150. doi:10.1107/S0907444905035365.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0015-834A-B Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0027-C55B-3
Genre: Journal Article

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1938168.pdf (Publisher version), 2MB
 
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 Creators:
Kettenberger, H., Author
Cramer, P.1, Author              
Affiliations:
1Department of Molecular Biology, MPI for Biophysical Chemistry, Max Planck Society, ou_1863498              

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Free keywords: multiprotein crystals; nucleic acids; fluorescence; soaking; DNA; RNA; cocrystallization; protein complexes; SYBR Gold.
 Abstract: A highly sensitive and specific assay for the rapid detection of nucleic acids in crystals of macromolecular complexes is reported. The assay is based on the observation of crystals under a fluorescence microscope after incubation with a fluorescent dye that specifically binds nucleic acids. The assay allows clear distinction between crystals of free RNA polymerase II and crystals of the polymerase bound to DNA and RNA or RNA alone, even when the nucleic acids account for only 3% of the crystalline material. Fluorescence detection was also used to monitor the diffusion and accumulation of a fluorescence-labelled polymerase-binding protein in preformed polymerase crystals. Additional experiments suggest the general applicability of these fluorescence assays for the characterization of crystal contents.

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Language(s): eng - English
 Dates: 2006-01-232006-02
 Publication Status: Published in print
 Pages: -
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 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1107/S0907444905035365
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Title: Acta Crystallographica Section D: Biological Crystallography
Source Genre: Journal
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Pages: - Volume / Issue: 62 (2) Sequence Number: - Start / End Page: 146 - 150 Identifier: -