Study on SNP Genotyping of Degraded DNA by Fluorescence-labeled Multiplex LDR-PCR Amplification
10.12007/j.issn.0258-4646.2017.08.008
- VernacularTitle: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
- Author:
Jiaxin XING
1
;
Yihua SUN
;
Jinfeng XUAN
;
Jun YAO
;
Mei DING
;
Hao PANG
;
Chunmei LI
;
Xi XIA
;
Baojie WANG
Author Information
1. 中国医科大学法医学院法医血清教研室
- Keywords:
forensics biological evidence;
ligase detection reaction;
compound amplification;
degraded DNA;
SNPs
- From:
Journal of China Medical University
2017;46(8):703-709
- CountryChina
- Language:Chinese
-
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.
