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The agar contact replica technique after isoelectric focusing as a screening method for the detection of enzyme variants
Author(s) -
Pretsch Walter,
Charles Daniel J.,
Narayanan Komaratchi R.
Publication year - 1982
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150030305
Subject(s) - isoelectric focusing , agar gel , agar , enzyme , polyacrylamide gel electrophoresis , chromatography , agar plate , replica , biology , electrophoresis , chemistry , biochemistry , microbiology and biotechnology , genetics , bacteria , art , visual arts
Electrofocusing has been adapted in order to make screening experiments for the detection of enzyme variants more efficient. The principle of the new technique consists in making agar copies of the polyacrylamide gel by contact; the original gel and the agar copies are then developed for different enzyme systems. By doing so, the number of enzymes, i.e. loci, per test animal screened after a single electrofocusing run is increased. This method has proved to be feasible for the detection of mouse variants with altered enzyme banding patterns due to parental treatment with chemical mutagens.