Cereal Chem 62:372-377   |  VIEW ARTICLE
Computer-Aided Analysis of Gliadin Electrophoregrams. I. Improvement of Precision of Relative Mobility Determination by Using a Three Reference Band Standardization.

H. D. Sapirstein and W. Bushuk. Copyright 1985 by the American Association of Cereal Chemists, Inc. 

Gliadin electrophoregrams used to identify wheat cultivars were analyzed by a computer-aided procedure. The reproducibility of relative mobility (Rm) determination using a standardized electrophoretic technique in 6% polyacrylamide gels was evaluated over a five-month period. Migration distances of bands were measured from photographs of electrophoregrams using a digitizing tablet that fed the values directly to a computer for processing. Absolute positional data was normalized to an Rm scale using the single reference band 50 nomenclature of Bushuk and Zillman (1978) and a new three reference band system. The variation in Rm increased with increasing distance between a particular band and the reference band(s). Compared to the single reference band procedure, the use of three reference bands significantly improved precision of results by stabilizing the variability in Rm measurements at a low level over the entire electrophoregram. Mean standard errors below 0.08 Rm units were achieved with minimal replication, effectively increasing the discriminative power of electrophoresis for comparative analysis.