Quantification of picogram levels of specific DNA immobilized in microtiter wells

A non-radioisotopic and sensitive method for quantification of specific DNA immobilized in microtiter wells has been developed. This method is based upon the immobilization of DNA in microtiter wells and hybridization with biotinylated DNA probe which is followed by complexing with avidin-β-galactos...

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Bibliographic Details
Published in:FEBS letters 1985-04, Vol.183 (2), p.379-382
Main Authors: Nagata, Yoshiho, Yokota, Hiroshi, Kosuda, Osamu, Yokoo, Keiko, Takemura, Kazuo, Kikuchi, Tetsuya
Format: Article
Language:English
Subjects:
Analytical, structural and metabolic biochemistry
Avidin
Avidin-biotinylated β-galactosidase complex
Bacteriophage lambda - genetics
beta-Galactosidase - metabolism
Biological and medical sciences
Biotin
DNA probe quantitation
Dna, deoxyribonucleoproteins
DNA, Viral - analysis
Fundamental and applied biological sciences. Psychology
Immobilized DNA
Microchemistry - methods
Microtiter well
Nowadioisotopic DNA probe
Nucleic Acid Hybridization
Nucleic acids
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Summary:A non-radioisotopic and sensitive method for quantification of specific DNA immobilized in microtiter wells has been developed. This method is based upon the immobilization of DNA in microtiter wells and hybridization with biotinylated DNA probe which is followed by complexing with avidin-β-galactosidase. By measuring fluorescence emitted from the hydrolyzed product by β-galactosidase of 4-methylumbelliferyl-β-D-galactoside, it has become possible to quantify a few picograms of specific DNA in DNA samples immobilized in plastic microtiter wells.
ISSN:0014-5793
1873-3468
DOI:10.1016/0014-5793(85)80814-0