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Sources of Variation in Computer Imaging of Electrophoretic Gels
Author(s) -
Bell G. E.,
McDonald M. B.,
Danneberger T. K.,
St. Martin S. K.
Publication year - 1998
Publication title -
crop science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.76
H-Index - 147
eISSN - 1435-0653
pISSN - 0011-183X
DOI - 10.2135/cropsci1998.0011183x003800020041x
Subject(s) - isoelectric focusing , electrophoresis , esterase , biology , variation (astronomy) , cultivar , biological system , chromatography , analytical chemistry (journal) , botany , chemistry , genetics , biochemistry , physics , astrophysics , enzyme
Electrophoresis is commonly used in agricultural sciences and industries to evaluate protein constituents of plant and animal tissue for cultivar or species identification. Visual analysis of polymorphic bands is effective, but comparison of electrophoretic banding patterns containing common bands of differing optical densities is difficult and subjective. A reliable and reproducible objective means of evaluation is needed. The purpose of this study was to test computer‐assisted image analysis for quantitative evaluation of electrophoretic bands. Seed protein was extracted from mixtures and blends of turfgrass species and cultivars, separated on isoelectric focusing gels, and stained for esterase activity. Banding patterns were scanned and computer‐assisted evaluation of band densities performed. Statistical evaluation of repetitive scans of varying band densities revealed that significant variation occurred among gels, scans,and among gel positions. However, when multiple bands from the same sources were averaged across a single gel and evaluated from a single scan, computer‐assisted image analysis prove accurate for discriminating bands with minor density variation. Computer‐assisted image analysis is an effective quantitative method for evaluation of electrophoretic gels when techniques are used to minimize variation.