1. A LDR-PCR Approach for Multiplex Polymorphisms Genotyping of Severely Degraded DNA with Fragment Sizes <100 bp
- Author
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Bao-jie Wang, Hong-Yu Guan, Hao Pang, Jin-Feng Xuan, and Zhen Zhang
- Subjects
Genotype ,Ligase Chain Reaction ,DNA Fragmentation ,Biology ,Polymerase Chain Reaction ,Polymorphism, Single Nucleotide ,Pathology and Forensic Medicine ,law.invention ,law ,Genetics ,Humans ,Multiplex ,Ligase chain reaction ,Genotyping ,Alleles ,Polymerase chain reaction ,DNA Primers ,chemistry.chemical_classification ,DNA ligase ,Hybridization probe ,DNA Degradation, Necrotic ,Amplicon ,DNA Fingerprinting ,Molecular biology ,chemistry ,DNA Probes - Abstract
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
- Published
- 2009