Application Research of DNATyper mtDNA-SNP60? Kit in Forensic DNA Casework
The DNATyper mtDNA-SNP60™ kit is a mtDNA SNP multiplex amplification system,which is suitable for the Chinese population.At present,the DNATyper mtDNA-SNP60™ kit has not been officially launched on the market.There has been no systematic research on the application of the DNATyper mtDNA-SNP60™ kit in practical cases.To explore the feasibility of the application of DNATyper mtDNA-SNP60™ kit in forensic DNA casework,DNATyper mtDNA-SNP60™ kit was used to test the mtDNA SNP of 100 unrelated individuals of Han nationality and 20 groups of whole siblings.25 pg/μL DNA samples from horses,cows,sheep,pigs,chickens,ducks,cats,dogs,rabbits,mice,and Escherichia coli were taken to perform species specific testing.5,10,20,40 μmol/L heme were taken for anti inhibitory testing.Two batches of DNATyper mtDNA-SNP60™ reagent kits were performed stability testing after repeated freeze-thaw for 10 times.VeriFiler™ Plus PCR amplification kit and DNATyper mtDNA-SNP60™ kit were used to detect the 100 old,rotten and degraded samples respectively.Results showed that all 100 unrelated individuals of Han nationality had clear mtDNA SNP typing,and the results were completely consistent with those obtained by mtDNA sequencing.Under the detection of DNAtyper mtDNA-SNP60™ kit,the 100 unrelated individuals of Han nationality showed 100 different haplotypes,and the mtDNA SNP typing was the same among individuals in each group of the 20 whole siblings'groups,which suggested that the DNATyper mtDNA-SNP60™ kit can be used for maternal lineage identification.DNATyper mtDNA-SNP60™ kit was used to detect DNA samples from horses,cows,sheep,pigs,chickens,ducks,cats,dogs,rabbits,mice,and Escherichia coli,but no specific typing was found,indicating that the kit has good species specificity.When heme concentration at ≤ 40 μmol/L,all mtDNA SNP sites were correctly typed,indicating that the kit has certain anti heme ability.Two batches of DNATyper mtDNA-SNP60™reagent kits were subjected to repeated freeze-thaw for 10 times,and all mtDNA SNP sites were correctly classified,indicating that the kit has good stability.Detection rate of STR for the 100 old,rotten and degraded samples was 55％,while the detection rate of mtDNA SNP was 86％,which was significantly higher than that of STR.When template DNA concentration was above 5 pg/μL,DNATyper mtDNA-SNP60™ kit would obtain a complete typing result.In conclusion,the DNATyper mtDNA-SNP60™ kit can achieve composite amplification of mtDNA SNP on existing STR testing platforms,and the results will be accurate and reliable.Besides,DNATyper mtDNA-SNP60™ kit can significantly improve the detection rate of aged,rotten,and degraded samples,and can be a useful supplementary means of STR examination.This study lays the foundation for the subsequent development of the DNATyper mtDNA-SNP60™ kit.
forensic geneticDNATyper mtDNA-SNP60™ kitmitochondrial DNAsingle nucleotide polymorphismsVeriFiler™ Plus PCR amplification kitshort tandem repeat
杨乐、陈滢、吴俞衡、石妍、齐朝阳、孔祥仕、马温华
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哈尔滨市公安局刑事技术支队,哈尔滨 150010
北京通达首诚司法鉴定所,北京 100192
北京市公安局房山分局,北京 102488
公安部鉴定中心,北京市现场物证检验工程技术研究中心,北京 100038
吉首大学旅游学院生态旅游湖南省重点实验室,湖南张家界 427000
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法医遗传学 DNATyper mtDNA-SNP60™试剂盒 线粒体DNA 单核苷酸多态性 VeriFiler™ Plus PCR扩增试剂盒 短串联重复序列
国家科技期刊平台
NSTL
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