Disadvantage of the in silico method. Genes or non-coding regions of interest have to be identified and sequenced before the technique can be used. Once images are obtained in a digitized form, they are transferred to a separate workstation for analysis. A variety of software programs are being developed to accomplish the analysis efficiently. Signals are measured as absolute intensities for a given target, or as ratios of two probes with different fluorescent labels representing two separate treatments to be compared or with one probe as an internal control.
Background is subtracted by different methods, depending on the software used.
The ratio of two signals provides relative response ratios rather than an estimate of an absolute signal. 2001 Ruth Alscher, All rights reserved.