title: Exploring of tri-allelic SNPs using pyrosequencing and the SNaPshot methods for forensic application.
abstract: Tri-allelic single nucleotide polymorphisms (SNPs) are potential forensic markers for DNA analysis. Currently, only a limited number of tri-allelic SNP loci have been proved to be fit for forensic application. In this study, we aimed to develop an effective method to select and genotype tri-allelic SNPs based on both Pyrosequencing (PSQ) and the SNaPshot methods. 50 candidate SNPs were chosen from NCBI's dbSNP database and were analyzed by PSQ. The results revealed that 20 SNPs were tri-allelic and were located on 16 autosomal chromosomes. Then 20 SNP loci were combined in one multiplex polymerase chain reaction to develop a single base extension (SBE)-based SNP-typing assay. A total of 100 unrelated Chinese individuals were genotyped by this assay and allele frequencies were estimated. The total discrimination power was 0.999999999975 and the cumulative probability of exclusion was 0.9937. These data demonstrated that the strategy is a rapid and effective method for seeking and typing tri-allelic SNPs. In addition, the 20 tri-allelic SNP multiplex typing assay may be used to supplement paternity testing and human identification.
authors.affiliation: Department of Forensic Genetics, School of Basic Science and Forensic Medicine, Sichuan University, Chengdu, P. R. China.
authors.forename: Lagabaiyila
authors.initials: L
authors.lastname: Zha
authors.forename: Libing
authors.initials: L
authors.lastname: Yun
authors.forename: Pengyu
authors.initials: P
authors.lastname: Chen
authors.forename: Haibo
authors.initials: H
authors.lastname: Luo
authors.forename: Jing
authors.initials: J
authors.lastname: Yan
authors.forename: Yiping
authors.initials: Y
authors.lastname: Hou
mesh_terms: D000483:Alleles; D002681:China; D044707:Forensic Sciences / Q000379:methods*; D005838:Genotype; D006801:Humans; D015810:Linkage Disequilibrium; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D017422:Sequence Analysis, DNA / Q000379:methods*