DNA sequencing using a four-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 transfer 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 parallel. The resulting four-color image files are automatically reduced to four-color line plots, and a base-calling program (Sax) is used to call the sequence. The performance of this system for DNA sequencing is demonstrated by examining twelve different motifs of the hypervariable region I of human mitochondrial (mt) DNA obtained from a Sierra Leone population.
Kheterpal
I
I
Department of Chemistry, School of Public Health, University of California, Berkeley 94720, USA.
Scherer
J R
JR
Clark
S M
SM
Radhakrishnan
A
A
Ju
J
J
Ginther
C L
CL
Sensabaugh
G F
GF
Mathies
R A
RA
D001483:Base Sequence; D004272:DNA, Mitochondrial / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000295:instrumentation* / Q000379:methods; D006801:Humans; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA / Q000295:instrumentation* / Q000379:methods; D012807:Sierra Leone
D004272:DNA, Mitochondrial