A multiple-capillary electrophoresis system for small-scale DNA sequencing and analysis.
A five-capillary system has been developed for DNA sequencing and analysis. The post-column fluorescence detector is based on a sheath-flow cuvette. The instrument provides uniform and continuous illumination of the samples. The cuvette virtually eliminates cross-talk in the fluorescence signal between capillaries. Discrete single-photon counting avalanche photodiodes provide high efficiency light detection. The instrument has detection limits (3sigma) of 130 +/- 30 fluorescein molecules injected onto each capillary. Over 650 bases of sequence at 98.8% accuracy were generated in 100 min at 50 degrees C from M13mp18. Separation and detection of short tandem repeats proved efficient and accurate with the use of internal standards for direct comparison of migration times between capillaries.
Zhang
J
J
Department of Chemistry, University of Alberta, Edmonton, Alberta T6G 2G2, Canada.
Voss
K O
KO
Shaw
D F
DF
Roos
K P
KP
Lewis
D F
DF
Yan
J
J
Jiang
R
R
Ren
H
H
Hou
J Y
JY
Fang
Y
Y
Puyang
X
X
Ahmadzadeh
H
H
Dovichi
N J
NJ
D000328:Adult; D002648:Child; D002897:Chromosomes, Human, Pair 7; D019075:Electrophoresis, Capillary / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D005260:Female; D005336:Fiber Optic Technology; D005453:Fluorescence; D006801:Humans; D008297:Male; D018895:Microsatellite Repeats; D008853:Microscopy; D055095:Optics and Photonics; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000295:instrumentation* / Q000379:methods; D020080:Tandem Repeat Sequences; D013696:Temperature