荧光染料掺入PCR检测STR基因座多态性的研究

Study of Fluorescent Dry Incorporation Technology to PCR - STR

目的研究荧光染料掺入PCR检测STR基因座多态性。方法 利用荧光染料掺人PCR扩增技术,测定FABP、CYP19、FOLP23、MBP、DYS19五个基因座的梯阶片段长度,将带有荧光标记物的dCTP通过PCR反应加入到目的片段PCR产物中,通过PE 377 DNA测序仪分离和基因扫描软件分析。结果 获得了清晰、准确的图谱,计算出各基因座中核心序列的重复次数,并按照ISFH标准对各基因座命名。结论 该方法具有灵敏度高、结果客观准确、操作简便、产物定量等优点,在新基因座开发和法医鉴定实践中具有较理想的应用价值和发展前景。

Objective Establishment of a new method for PCR products length measurement by fluorescent dye incorporation technology. Methods Using fluorescent dye labeled dCTP, we incorporated fluorescent signal to PCR products of 5 STR loci: FABP, CYP19, FOLP23, MBP, DYS19. And the length of products was analyzed by collection software and Genescan software through PE 377 DNA sequencer. Results The fragment length data of every alleles of 5 loci described above was obtained and the value of fragment length was transferred to the repeats number of these loci so that alleles were nominated to certain numbers under the ISFH nomenclature criteria. Conclusions Advantage of method estabolished in our research is highly sensitive, simple and convenient, accurate, relatively quantitative and is very applicable and valuable to research work and case investigation.