Detection of amplified Y chromosome-specific sequence by capillary electrophoresis with laser-induced fluorescence.
OBJECTIVE
To develop a sensitive method for genetic diagnosis using capillary electrophoresis with laser-induced fluorescence.


METHODS
Using male DNA diluted with female DNA as an example, ZFY gene from Y chromosome was amplified specifically by polymerase chain reaction (PCR) with fluorescence-labeled primers and detected by capillary electrophoresis with laser-induced fluorescence.


METHODS
Laser operating laboratory.


METHODS
Human male and female DNA were extracted from healthy human male and female subjects.


METHODS
None.


METHODS
The concentration of human DNA was determined by using a spectrophotometer at an absorbance of 260 nm.


RESULTS
Deoxyribonucleic acid fragments amplified from a single copy of ZFY gene were detected by capillary electrophoresis with laser-induced fluorescence after 35 cycles of PCR amplification.


CONCLUSIONS
This method is potentially applicable for rapid and sensitive detection of fetal Y chromosome DNA sequence in maternal circulation and of single-cell DNA diagnosis.
Liu
M S
MS
Advanced Technology Center, Beckman Instruments Inc., Fullerton, California 92634, USA.
Rampal
S
S
Evangelista
R A
RA
Lee
G C
GC
Chen
F T
FT
D001483:Base Sequence; D004247:DNA / Q000737:chemistry*; D004586:Electrophoresis; D005260:Female; D005453:Fluorescence; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D014998:Y Chromosome
D004247:DNA