Premium
Technical improvements in two‐dimensional electrophoresis increase the level of genetic variation detected in wheat‐seedling proteins
Author(s) -
Damerval Catherine,
De Vienne Dominique,
Zivy Michel,
Thiellement Hervé
Publication year - 1986
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.1150070108
Subject(s) - isoelectric focusing , chemistry , chromatography , trichloroacetic acid , electrophoresis , sodium dodecyl sulfate , extraction (chemistry) , seedling , acetone , gel electrophoresis , genetic variation , sodium carbonate , potassium carbonate , urea , sodium , biochemistry , botany , biology , gene , organic chemistry , enzyme
The level of genetic variation revealed by two‐dimensional electrophoresis of proteins from seedlings of two wheat lines strongly depends on the technical procedures. Improvements in extraction and electrophoresis procedures relative to earlier experiments on the same material led to a significant increase in the genetic variation revealed: 15.2 % instead of 6.7 % of the spots were genetically variable. The improved procedure is based on (i) precipipation of proteins from wheat seedlings with trichloroacetic acid and acetone, (ii) solubilization of the proteins with a solution containing urea, potassium carbonate and sodium dodecyl sulfate, (iii) isoelectric focusing in an optimized pH gradient, obtained with a mixture of carrier ampholytes (Pharmalyte and Servalyt), and (iv) running elecrophoresis in the second dimension on gels with increased surface.