With the recent rapid expansion in the use of the comparative genomic
hybridization (CGH) technique, increased attention to quality control
is essential, In the present study, we show that despite optimization
and standardization of metaphase preparation techniques and the commer
cial availability of metaphase spreads, batch-to-batch variability of
the preparations remains a significant problem. To facilitate reliable
CGH analysis despite this variability, we have developed a rapid dena
turation test to assess the quality of the preparations without hybrid
ization and quantitative image analysis criteria for assuring the day-
to-day quality of CGH experiments, including sensitivity, specificity,
and dynamic range, Monitoring the dynamic range of the hybridizations
was found to be particularly critical for achieving sensitive and rel
iable CGH results. This reliability can be achieved, for example, by h
ybridization of a green-labeled normal male DNA against red-labeled fe
male DNA and monitoring of the green:red ratio of the X chromosome in
relation to that of the autosomes. (C) 1997 Wiley-Liss, Inc.