国产磁珠用于法医DNA检验的研究

发布时间：2018-04-13 03:15

本文选题：法医DNA检验 + 国产磁珠法　；参考：《东北师范大学》2009年硕士论文

【摘要】： 磁珠法实用范围广,可以用于血斑、唾液斑、精斑、汗斑等大多数生物样本的提取,方法操作简单,不需要特殊的分离设备,同时也避免了大样品体系需要反复离心而造成的DNA损失。本实验利用一款国产磁珠,在DNA回收纯化方面进行了一些探索。实验证明,国产磁珠用于DNA回收纯化具有高回收灵敏度、纯化效果好的特性,适合在案件检验中应用。首先,用国产磁珠法、硅珠法和IQ试剂盒分别对不同量的9947标准DNA进行提取,使用PowerPlex16试剂盒、ABI9700型扩增仪进行PCR复合扩增,扩增产物应用ABI-3100型DNA分析仪检测。实验表明,国产磁珠法在荧光定量PCR方面要优于硅珠法和IQ试剂盒。然后,用国产磁珠法、硅珠法和IQ试剂盒分别对稀释30倍、60倍的血液样本进行提取,使用PowerPlex16试剂盒、ABI9700型扩增仪进行PCR复合扩增,扩增产物应用ABI-3100型DNA分析仪检测。结果表明,国产磁珠法在痕量DNA提取方面要优于硅珠法和IQ试剂盒。最后,用国产磁珠法分别对三种污染检材:带血迹玻璃、衣领脱落细胞、牙刷进行提取,使用PowerPlex16试剂盒、ABI9700型扩增仪进行PCR复合扩增,扩增产物应用ABI-3100型DNA分析仪检测,16个STR基因座均获得了理想的DNA分型。结论,国产磁珠法能够从含有染料、血红素、金属离子等污染成分的样本中回收纯化DNA,并适合复合STR扩增。
[Abstract]:The magnetic bead method has a wide range of applications. It can be used to extract most biological samples such as blood spot, saliva spot, sperm spot, sweat spot and so on. The method is simple to operate and does not require special separation equipment.At the same time, the DNA loss caused by repeated centrifugation of large sample system is avoided.In this experiment, a domestic magnetic beads were used to recover and purify DNA.It is proved by experiments that the domestic magnetic beads have high recovery sensitivity and good purification effect for DNA recovery and purification, and are suitable for application in case testing.Firstly, the 9947 standard DNA was extracted by magnetic bead method, silicon bead method and IQ kit respectively. The PCR was amplified by PowerPlex16 kit ABI9700 amplification instrument, and the amplification product was detected by ABI-3100 DNA analyzer.The results show that the domestic magnetic bead method is superior to the silicon bead method and IQ kit in fluorescence quantitative PCR.Then, the blood samples with 30 times dilution and 60 times dilution were extracted by magnetic bead method, silicon bead method and IQ kit respectively. The PCR was amplified by PowerPlex16 kit ABI9700, and the products were detected by ABI-3100 DNA analyzer.The results showed that the domestic magnetic bead method was superior to the silicon bead method and IQ kit in the extraction of trace DNA.Finally, three kinds of contaminated samples were extracted by domestic magnetic bead method: stained glass, neckline exfoliated cells, toothbrush, and PCR was amplified by PowerPlex16 kit ABI9700.The amplification products were detected by ABI-3100 type DNA analyzer, and all 16 STR loci obtained ideal DNA typing.Conclusion: the domestic magnetic beads method can recover and purify DNA from samples containing dye, heme and metal ions, and is suitable for compound STR amplification.
【学位授予单位】：东北师范大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：D919

【引证文献】