DNA-SEQUENCING USING A 4-COLOR CONFOCAL FLUORESCENCE CAPILLARY ARRAY SCANNER

The design, construction and operation of a four-color capillary array electrophoresis scanner are presented. The use of sensitive energy tr ansfer primers facilitates four-color detection of the DNA sequencing fragments following excitation at a single laser wavelength (488 nm). This scanner collects fluorescence data from up to 25 capillaries in p arallel. The resulting four-color image files are automatically reduce d to four-color line plots, and a base-calling program (Sax) is used t o cal the sequence. The performance of this system for DNA sequencing is demonstrated by examining twelve different motifs of the hypervaria ble region I of human mitochondrial (mt) DNA obtained from a Sierra Le one population.