Abstract 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.
A method for preparing an electrophoresis gel medium while minimizing exposure to harmful chemicals which comprises forming a solution of the copolymer of a kit for preparing electrophoresis gel media which comprises, in combination, (1) a copolymer consisting of poly[acrylamide-co-N-(3-acetoacetaamidopropyl]methacrylamide having a weight ratio of acrylamide to the comonomer of 95:5 and (2) in a package separate from said copolymer, a crosslinking agent for reacting with the functional groups on the repeating units in the said copolymer derived from a monomer that contains a functional group that will enter into a crosslinking reaction by other than a free-radical initiated mechanism and an electrophoresis buffer in deionized water employing such proportions of said copolymer, buffer and water as to provide a gel of the desired said copolymer concentration and pH, providing means with which to form a shaped electrophoresis gel medium of the desired dimensions, adding the crosslinking agent from the kit to said solution of said copolymer and buffer in such concentration of crosslinking agent as to cause gelation to occur within a time period of from about 5 minutes to about 15 hours after said addition, and then promptly employing said gel shaping means to form the gel medium of desired dimensions from the gel thereby produced.
Claim 2
Independent Claim
208 words
A method for preparing an electrophoresis gel medium while minimizing exposure to harmful chemicals which comprises forming a solution of the copolymer of a kit for preparing electrophoresis gel media which comprises, in combination, (1) a copolymer consisting of poly[acrylamide-co-N-(3-acetoacetaamidopropyl]methacrylamide having a weight ratio of acrylamide to the comonomer of 95:5 and (2) in a package separate from said copolymer, a crosslinking agent for reacting with the functional groups on the repeating units in the said copolymer derived from a monomer that contains a functional group that will enter into a crosslinking reaction by other than a free-radical initiated mechanism and an electrophoresis buffer in deionized water employing such proportions of said copolymer, buffer and water as to provide a gel of the desired said copolymer concentration and pH, providing means with which to form a shaped electrophoresis gel medium of the desired dimensions, adding the crosslinking agent from the kit to said solution of said copolymer and buffer in such concentration of crosslinking agent as to cause gelation to occur within a time period of from about 5 minutes to about 15 hours after said addition, and then promptly employing said gel shaping means to form the gel medium of desired dimensions from the gel thereby produced.
Claim 3
Dependent Claim
10 words
The method of claim 1 wherein said buffer is tris(hydroxymethyl)aminoethane.
Claim 4
Dependent Claim
11 words
The method of claim 1 wherein said crosslinking agent is glyoxal.