DETECTION AND QUANTITATION OF UNLABELED NUCLEIC-ACIDS IN POLYACRYLAMIDE GELS

Authors
Citation
P. Hendry et G. Hannan, DETECTION AND QUANTITATION OF UNLABELED NUCLEIC-ACIDS IN POLYACRYLAMIDE GELS, BioTechniques, 20(2), 1996, pp. 258-264
Citations number
3
Categorie Soggetti
Biochemical Research Methods
Journal title
ISSN journal
07366205
Volume
20
Issue
2
Year of publication
1996
Pages
258 - 264
Database
ISI
SICI code
0736-6205(1996)20:2<258:DAQOUN>2.0.ZU;2-3
Abstract
A method for the detection and quantitation of unlabeled nucleic acids in polyacrylamide gels is presented. The technique is a variation oil UV shadowing. The technique is based on the observation that some sto rage phosphor screens (Kodak in this case) are sensitive to UV light i n the range where nucleic acids absorb most strongly. Nucleic acid pre sent in a gel resting on the screen absorbs the UV light and casts an ultraviolet shadow on the storage phosphor screen. The amount of nucle ic acid in a particular area of the polyacrylamide gel is inversely pr oportional to the average pixel intensity in the corresponding area of the image. The technique therefore provides an electronic record of t he gel in addition to the ability to quantitate the amount of nucleic acid present in any particular band. The storage phosphor screen image s are erased by visible light. If the passage of that Eight to the scr een is blocked by the presence of solid objects or images, the reverse process can be achieved, i.e., the recording of positive images by vi sible Eight etching of a uniformly charged background By this method, we are able to transform old autoradiographic data into digital form f or easy storage, computerized manipulation and distribution.