Study on Optimal Conditions in Arbitrarily Primed PCR Human DNA Fingerprinting
- VernacularTitle:随机引物PCR检测人DNA指纹条件的优化
- Author:
Dayue TONG
;
Ping XU
;
Yubin GUO
;
Fang LI
;
Da LIN
;
Jinghua OU
;
Xinyao WU
- Publication Type:Journal Article
- From:
Journal of Sun Yat-sen University(Medical Sciences)
2001;22(3):231-234
- CountryChina
- Language:Chinese
-
Abstract:
【Objective】To explore the optimal conditions in fingerprinting (APHDF).【Methods】The human DNA fingerprints were detected by APHDF.A pair of short primers was used for amplification.The experimental conditions including template,Mg2+,deoxyribonucleotides,and parameters of cycle,were optimized.【Results】The template DNA should be abstracted freshly and the concentration should be ranged from 50～550 mg/L.The best concentration of Mg2+was 5.0 mmol/L.The deoxyribonucleotides concentration was optimal at 0.2 mmol/L.The PCR cycling parameters were as follows :The denaturing temperatures,annealing temperatures and extension temperatures were 94 ℃ and 90 ℃ for 30 s,43 ℃ and 48 ℃ for 40 s or 50 s,and 72 ℃ for 1 min or 80 s,respectively.【Conclusion】The optimal conditions of the experiment are obtained,with good reproducibility and high specificity.Therefore,this method can be widely applied in practice.
