Zhang L, Xin J, Chen G, Li R, Chen H
College of Forensic Medicine, WCUMS, Chengdu 610041, China.
Hua Xi Yi Ke Da Xue Xue Bao. 2001 Mar;32(1):5-8.
We have designed a new method to produce standard DYS385 allelic ladder in order to solve the problem of the accuracy and standardization of STR-PCR typing in forensic practice.
Nine different PCR amplified DYS385 allelic fragments were isolated from the gel, eluted into the distilled water and reamplified by PCR. The purified allelic fragments were then blunt-end subcloned individually into the pUC plasmid vectors and transfected into competent E. coli DH5 alpha cells. The sequencing results confirmed that the size and the constructure of the inserts were correct. The recombinant plasmids DNA with 9 inserts were then used as templates for reamplification to generate DYS385 standard ladder.
A large quantity of standard DYS385 allelic ladder was obtained, with which the genetic polymorphisms of DYS385 locus both in Chinese Han and German populations were studied.
The STR standard ladder constructed by this method has high value in the forensic application, and the DYS385 locus is robust for forensic analysis.
设计一种新方法制备标准DYS385等位基因阶梯，以解决法医实践中STR-PCR分型的准确性和标准化问题。
从凝胶中分离出9个不同的PCR扩增DYS385等位基因片段，洗脱到蒸馏水中，再通过PCR重新扩增。然后将纯化的等位基因片段分别平端亚克隆到pUC质粒载体中，并转入感受态大肠杆菌DH5α细胞。测序结果证实插入片段的大小和结构正确。然后将带有9个插入片段的重组质粒DNA用作重新扩增的模板，以生成DYS385标准阶梯。
获得了大量标准DYS385等位基因阶梯，并用其研究了中国汉族和德国人群中DYS385基因座的遗传多态性。
该方法构建的STR标准阶梯在法医应用中具有很高的价值，DYS385基因座在法医分析中表现稳定。
