high-capacity column for affinity purification of sequence-specific DNA-binding proteins

We report an improved method for synthesis of a DNA affinity column, involving site-specific linkage of a monomeric or concatenated oligonucleotide to an activated chromatographic support. In the present method, efficient and site-specific coupling is accomplished between DNA bearing a reactive alky...

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Bibliographic Details
Published in:Nucleic acids research 1992-07, Vol.20 (13), p.3525-3525
Main Authors: Larson, C.J, Verdine, G.L
Format: Article
Language:English
Subjects:
Adenine - analogs & derivatives
Analytical, structural and metabolic biochemistry
Base Sequence
Biological and medical sciences
Chromatography, Affinity - instrumentation
DNA-binding proteins
DNA-Binding Proteins - isolation & purification
DNA-Binding Proteins - metabolism
Fundamental and applied biological sciences. Psychology
General aspects, investigation methods
Molecular Sequence Data
Oligodeoxyribonucleotides - genetics
Oligodeoxyribonucleotides - metabolism
Phosphorylation
Proteins
purification
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Summary:We report an improved method for synthesis of a DNA affinity column, involving site-specific linkage of a monomeric or concatenated oligonucleotide to an activated chromatographic support. In the present method, efficient and site-specific coupling is accomplished between DNA bearing a reactive alkylamine-tethered nucleoside and a carboxyl-activated support; attachment of the tether to a DNA base (6-8) and co-synthetic 5'-phosphorylation allow for facile generation of concatenated DNA.
ISSN:0305-1048
1362-4962
DOI:10.1093/nar/20.13.3525