中山医科大学学报2001，Vol.22Issue(3)：231-234,4.
随机引物PCR检测人DNA指纹条件的优化
Study on Optimal Conditions in Arbitrarily Primed PCR Human DNA Fingerprinting
童大跃 1徐平 1郭语彬 1李芳 1林达 1区敬华 2伍新尧2
作者信息
- 1. 中山医科大学生物化学教研室,
- 2. 中山医科大学法医学系,
- 折叠
摘要
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.关键词
AP-PCR/DNA指纹法分类
生物科学引用本文复制引用
童大跃,徐平,郭语彬,李芳,林达,区敬华,伍新尧..随机引物PCR检测人DNA指纹条件的优化[J].中山医科大学学报,2001,22(3):231-234,4.
