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目的研究GoldeneyeTM 20A-M试剂盒在DNA数据库建设中的应用。方法针对血滤纸特性,优化不同条件血痕样品的取样量、扩增循环次数,缩短PCR扩增时间,以建立适合批量样品直接快速扩增检验的方法,并将其应用于1 200份数据库样品的检验中。结果确定了批量样本检验的最适取样量和扩增循环次数,建立了快速扩增反应程序,1 200份样品的直接快速扩增检验成功率为98.4%,批量检验92份样品(1板)从取样到电泳完成只需6h。结论 GoldeneyeTM 20A-M试剂盒的应用,免去DNA提取过程,方法简单、快速、获得信息量大,适用于DNA数据库建设的需要。
Objective To study the application of GoldeneyeTM 20A-M in DNA database construction. Methods According to the characteristics of blood filter paper, the sampling volume of bloodstain samples in different conditions, the number of amplification cycles and the PCR amplification time were optimized to establish a direct rapid amplification test for batch samples and applied it to samples of 1200 samples Inspection. Results The optimum sampling volume and number of cycles for amplification were determined. A rapid amplification reaction program was established. The success rate of direct rapid amplification test of 1 200 samples was 98.4% and that of 92 samples (1 plate) Just 6h from sampling to electrophoresis. Conclusion The application of GoldeneyeTM 20A-M kit eliminates the need for DNA extraction. The method is simple, rapid and has a large amount of information. It is suitable for the construction of DNA database.