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Detection of amplified Y chromosome-specific sequence by capillary electrophoresis with laser-induced fluorescence
M S Liu1, S Rampal, R A Evangelista
1Advanced Technology Center, Beckman Instruments Inc., Fullerton, California 92634, USA.
Objective:
To develop a sensitive method for genetic diagnosis using capillary electrophoresis with laser-induced fluorescence.
Design:
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.
Setting:
Laser operating laboratory.
Participants:
Human male and female DNA were extracted from healthy human male and female subjects.
Interventions:
None.
Main Outcome Measures:
The concentration of human DNA was determined by using a spectrophotometer at an absorbance of 260 nm.
Result:
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.
Conclusion:
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.