Continuous, on-line DNA sequencing using a versatile infrared laser scanner/electrophoresis apparatus.
A new apparatus for continuously detecting fluorescently labeled DNA fragments is based on infrared fluorescence technology. This technology combines state-of-the-art developments in chemistry, laser technology, and detection, while achieving improved reliability, sensitivity, and flexibility for applications including DNA sequencing. DNA molecules labeled with a novel infrared fluorophore are detected during electrophoresis using a scanning infrared fluorescence microscope. The microscope consists of a laser diode for exciting the fluorophore and a silicon avalanche photodiode for detecting the infrared emission. Optimum conditions for detection and throughput are obtained by adjusting electrophoresis, scanning and imaging parameters. Typical DNA sequencing runs (test templates) allow identification of over 500 bases per sample with greater than 99% accuracy.
Middendorf
L R
LR
LI-COR, Inc., Lincoln, NE 68504.
Bruce
J C
JC
Bruce
R C
RC
Eckles
R D
RD
Grone
D L
DL
Roemer
S C
SC
Sloniker
G D
GD
Steffens
D L
DL
Sutter
S L
SL
Brumbaugh
J A
JA
D001483:Base Sequence; D004247:DNA / Q000737:chemistry*; D004277:DNA, Single-Stranded / Q000138:chemical synthesis; D004591:Electrophoresis, Polyacrylamide Gel / Q000295:instrumentation*; D007091:Image Processing, Computer-Assisted; D007834:Lasers; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D015394:Molecular Structure; D012815:Signal Processing, Computer-Assisted; D013055:Spectrophotometry, Infrared / Q000295:instrumentation; D013698:Templates, Genetic
D004277:DNA, Single-Stranded; D004247:DNA