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A thin‐layer multistrip polyacrylamide gel electrophoresis apparatus for ferguson plot analysis at the sub‐microgram load level
Author(s) -
Orbán László,
Sullivan James V.,
Zwieb Christian,
Chrambach Andreas
Publication year - 1989
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.1150101013
Subject(s) - polyacrylamide , polymerization , chromatography , materials science , layer (electronics) , analytical chemistry (journal) , viscosity , agarose , gel electrophoresis , chemistry , composite material , polymer , polymer chemistry , biochemistry
A procedure was developed for casting thin‐layer multistrip polyacrylamide gels and using them for the simultaneous gel electrophoresis at several gel concentrations (Ferguson plot analysis) at the sub‐microgram load level, using silver staining, autoradiography and, potentially, blotting for detection. The lower viscosity of polymerization mixtures, compared to agarose gelation mixtures, required the redesign of the multistrip cassette with separation of channels by rubber gaskets and the application of a cassette press. The lowered viscosity also required addition of 35% sucrose and an increased rate of polymerization in application to multistrip gels formed on a common NetFix backing. The present design allows one to obtain Ferguson plots exemplified by those of 32 P‐labeled DNA followed by autoradiographic detection.