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A LDR‐PCR Approach for Multiplex Polymorphisms Genotyping of Severely Degraded DNA with Fragment Sizes <100 bp *
Author(s) -
Zhang Zhen,
Wang BaoJie,
Guan HongYu,
Pang Hao,
Xuan JinFeng
Publication year - 2009
Publication title -
journal of forensic sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.715
H-Index - 96
eISSN - 1556-4029
pISSN - 0022-1198
DOI - 10.1111/j.1556-4029.2009.01166.x
Subject(s) - amplicon , genotyping , biology , polymerase chain reaction , multiplex , multiplex polymerase chain reaction , microbiology and biotechnology , genetics , dna , str analysis , sequencing by ligation , in silico pcr , typing , primer dimer , multiplex ligation dependent probe amplification , variants of pcr , str multiplex system , microsatellite , dna ligase , allele , genotype , gene , exon , genomic library , base sequence
Reducing amplicon sizes has become a major strategy for analyzing degraded DNA typical of forensic samples. However, amplicon sizes in current mini‐short tandem repeat‐polymerase chain reaction (PCR) and mini‐sequencing assays are still not suitable for analysis of severely degraded DNA. In this study, we present a multiplex typing method that couples ligase detection reaction with PCR that can be used to identify single nucleotide polymorphisms and small‐scale insertion/deletions in a sample of severely fragmented DNA. This method adopts thermostable ligation for allele discrimination and subsequent PCR for signal enhancement. In this study, four polymorphic loci were used to assess the ability of this technique to discriminate alleles in an artificially degraded sample of DNA with fragment sizes <100 bp. Our results showed clear allelic discrimination of single or multiple loci, suggesting that this method might aid in the analysis of extremely degraded samples in which allelic drop out of larger fragments is observed.