Document Detail


Mica functionalization for imaging of DNA and protein-DNA complexes with atomic force microscopy.
MedLine Citation:
PMID:  23027008     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Surface preparation is a key step for reliable and reproducible imaging of DNA and protein-DNA complexes with atomic force microscopy (AFM). This article describes the approaches for chemical functionalization of the mica surface. One approach utilizes 3-aminopropyl-trietoxy silane (APTES), enabling one to obtain a smooth surface termed AP-mica. This surface binds nucleic acids and nucleoprotein complexes in a wide range of ionic strengths, in the absence of divalent cations and in a broad range of pH. Another method utilizes aminopropyl silatrane (APS) to yield an APS-mica surface. The advantage of APS-mica compared with AP-mica is the ability to obtain reliable and reproducible time-lapse images in aqueous solutions. The chapter describes the methodologies for the preparation of AP-mica and APS-mica surfaces and the preparation of samples for AFM imaging. The protocol for synthesis and purification of APS is also provided. The applications are illustrated with a number of examples.
Authors:
Luda S Shlyakhtenko; Alexander A Gall; Yuri L Lyubchenko
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  931     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2013  
Date Detail:
Created Date:  2012-10-02     Completed Date:  2013-02-11     Revised Date:  2013-12-10    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  295-312     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Aluminum Silicates / chemistry*
Bicyclo Compounds, Heterocyclic / chemical synthesis,  chemistry
DNA-Binding Proteins / chemistry*,  ultrastructure
Microscopy, Atomic Force / methods*
Nucleosomes / chemistry*,  ultrastructure
Organosilicon Compounds / chemical synthesis,  chemistry
Plasmids / chemistry,  ultrastructure
Silanes / chemistry
Surface Properties
Time-Lapse Imaging
Grant Support
ID/Acronym/Agency:
1P01GM091743-01A1/GM/NIGMS NIH HHS; 1R01 GM096039/GM/NIGMS NIH HHS; 1R01 GM096039-01A1/GM/NIGMS NIH HHS; P01 GM091743/GM/NIGMS NIH HHS; P01 GM091743/GM/NIGMS NIH HHS; R01 GM096039/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Aluminum Silicates; 0/Bicyclo Compounds, Heterocyclic; 0/DNA-Binding Proteins; 0/Nucleosomes; 0/Organosilicon Compounds; 0/Silanes; 0/amino-propyl-triethoxysilane; 12001-26-2/mica; 283-60-3/silatrane
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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