Eastman-kodak Company

Johnson & Johnson Clinical Diagnostics, Inc.

Kenneth G Christy, Jr

Ignazio S Ponticello

Hans W Osterhoudt

Edward R Scalice

John L Daiss

Theodore W Esders

David J Sharkey

Wayne A Bowman

Jon N Eikenberry

David B LaTart

Antibodies which are specific to a thermostable DNA polymerase can be used to reduce or eliminate the formation of non-specific products in polymerase chain reaction methods. These antibodies and other temperature sensitive inhibitors are effective to inhibit DNA polymerase enzymatic activity at a certain temperature T.sub.1 which is generally below about 85.degree. C. The inhibitors are irreversibly inactivated at temperature T.sub.2 which is generally above about 40.degree. C. T.sub.2 is also greater than T.sub.1. Such inhibitors can be supplied individually or in admixture with the DNA polymerase in a diagnostic test kit suitable for PCR.

Thermostable polymerase specific antibody-containing DNA amplification

DNA amplification with thermostable DNA polymerase and polymerase

Preformed, water-soluble, acrylamide based copolymers comprising a minor proportion of a comonomer, which comonomer contains a site for a crosslinking reaction with a selected crosslinking agent by a reaction that does not involve a free-radical vinyl addition mechanism, which copolymers have M.sub.w and M.sub.n values within selected ranges, can be conveniently and safely used to prepare electrophoresis gel media in situ.

Polymeric electrophoresis media

A polyacrylamide type gel medium for electrophoresis is conveniently and safely prepared prior to use by crosslinking a water-soluble copolymer of an acrylamide monomer and at least one comonomer copolymerizable therewith, which comonomer contains an active methylene group that will enter into a crosslinking reaction that does not involve a free radical vinyl addition mechanism.

Electrophoresis media comprising active methylene groups

A polyacrylamide type gel medium for electrophoresis is conveniently and safely prepared prior to use from a kit which comprises (1) a water-soluble copolymer of an acrylamide monomer and at least one comonomer copolymerizable therewith, which comonomer contains functional groups that permit crosslinking of said copolymer by a covalent but non-free radical initiated mechanism; and (2) a selected crosslinking agent in a package that separates it from the copolymer. Optionally, additional materials that may be usefully incorporated in the gel may be included in the kit, for example, a selected electrophoresis buffer and a complexing agent, such as SDS. The kit is used to prepare the gel by forming a solution of the copolymer in deionized water at the desired concentration of copolymer, adding a sufficient amount of any optional ingredients to provide the desired concentrations thereof, adding such concentration of the crosslinking agent as will provide a gelation time of between about 5 minutes and about 15 hours, and pouring the so prepared reaction mixture into a preselected mold or casting device or onto a support so as to form the resulting gel into the desired shape and size.

Kit for electrophoresis gel medium

Growing community of inventors

Rochester, NY, United States of America

Kenneth G Christy, Jr