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Clinical Chemistry 31: 1438-1443, 1985;
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Clinical Chemistry, Vol 31, 1438-1443, Copyright © 1985 by American Association for Clinical Chemistry

Use of DNA immobilized on plastic and agarose supports to detect DNA by sandwich hybridization

R Polsky-Cynkin, GH Parsons, L Allerdt, G Landes, G Davis and A Rashtchian

Cloned Salmonella DNA, which has been immobilized irreversibly on plastic and agarose solid supports, can form hybrids in both single- layer and "sandwich" hybridization protocols. In single-layer hybridization, 3 micrograms of immobilized DNA bound at least 30 fmol of a specific 800-base DNA sequence (equivalent to 8.5 ng, or the amount of that sequence present in 4 X 10(10) organisms). In a 4-h sandwich hybridization protocol, as little as 14 amol (equivalent to 8 pg, or the amount of that sequence present in 1 X 10(7) organisms) of a 1600-base sequence of DNA could be detected. The methods described should be applicable to use with any set of probes--not just from Salmonella--that fulfill the criteria specified. The ability to perform DNA hybridizations on solid-phase matrices such as those used for immunoassay should bring DNA hybridization into the realm of routine clinical laboratory procedures.




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Copyright © 1985 by the American Association for Clinical Chemistry.