中国医科大学学报2017，Vol.46Issue(8)：703-709,7.DOI:10.12007/j.issn.0258-4646.2017.08.008
荧光标记LDR-PCR复合扩增方法对高度降解DNA检材的SNPs分型研究
Study on SNP Genotyping of Degraded DNA by Fluorescence-labeled Multiplex LDR-PCR Amplification
摘要
Abstract
Objective In this study,a multiplex PCR amplification system was constructed based on fluorescent labeling PCR and LDR,to provide a new strategy for analyzing severely degraded DNA.Methods Eight SNP loci (rs10802248,rs10516197,rs10488372,rs2278945,rs4757318,rs4887255,rs4889002,and rs9304473) were selected.Their LDR probes and PCR primers of linked products were designed and synthesized.Ligase detection reaction,PCR amplification,and capillary gel electrophoresis (CEG) were performed to establish the multiplex LDR-PCR amplification system.Results The genotypes of these 8 loci were obtained simultaneously by the fluorescence-labeled multiplex LDR-PCR amplification method.The loci profiles obtained by fluorescence-labeled multiplex LDR-PCR amplification were in accordance with those obtained by direct sequencing of the polymorphic regions in samples from all individuals.By fluorescence-labeled multiplex LDR-PCR amplification,the 8 SNP loci were efficiently amplified from the severely degraded FFPET DNA.Conclusion Eight SNP loci results could be obtained simultaneously by using the multiplex LDR-PCR amplification system,which is a simple,efficient,and practical SNP genotyping method with accurate and reliable results for highly degraded samples.关键词
法医物证学/连接酶检测反应/复合扩增/降解DNA/单核苷酸多态性Key words
forensics biological evidence/ligase detection reaction/compound amplification/degraded DNA/SNPs分类
医药卫生引用本文复制引用
邢佳鑫,孙溢华,宣金锋,姚军,丁梅,庞灏,李春梅,夏皙,王保捷..荧光标记LDR-PCR复合扩增方法对高度降解DNA检材的SNPs分型研究[J].中国医科大学学报,2017,46(8):703-709,7.基金项目
辽宁省教育厅科学研究一般项目(L2013319) （L2013319）
