AUTOMATIC MATRIX DETERMINATION IN 4 DYE FLUORESCENCE-BASED DNA-SEQUENCING

The four dye fluorescence detection strategy is a widely used approach to automated DNA sequence analysis. An important aspect of data proce ssing in this approach is the multicomponent analysis to deduce the co ncentrations of four fluorophores from fluorescence emission intensiti es at four different wavelengths. This requires knowledge of the corre ct transformation matrix M. The matrix M is a function both of the flu orophores employed and the fluorescence detection system. M is typical ly determined either by a calibration process with individual dyes, or by choosing four well-separated individual peaks corresponding to the four different dyes. Both are time-consuming and complicated procedur es for routine use. An automatic scheme for finding M directly from ra w sequence data is presented here. This facilitates data analysis and the underlying algorithm may also find utility in other multispectral applications.