Use of DNA immobilized on plastic and agarose supports to detect DNA by sandwich hybridization.
Open Access
- 1 September 1985
- journal article
- research article
- Published by Oxford University Press (OUP) in Clinical Chemistry
- Vol. 31 (9) , 1438-1443
- https://doi.org/10.1093/clinchem/31.9.1438
Abstract
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.Keywords
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