Supported by a grant from the National Natural Science Foundation of China (No 30772460).
A LDR-PCR Approach for Multiplex Polymorphisms Genotyping of Severely Degraded DNA with Fragment Sizes <100 bp*
Article first published online: 5 OCT 2009
© 2009 American Academy of Forensic Sciences
Journal of Forensic Sciences
Volume 54, Issue 6, pages 1304–1309, November 2009
How to Cite
Zhang, Z., Wang, B.-J., Guan, H.-Y., Pang, H. and Xuan, J.-F. (2009), A LDR-PCR Approach for Multiplex Polymorphisms Genotyping of Severely Degraded DNA with Fragment Sizes <100 bp. Journal of Forensic Sciences, 54: 1304–1309. doi: 10.1111/j.1556-4029.2009.01166.x
- Issue published online: 23 OCT 2009
- Article first published online: 5 OCT 2009
- Received 18 June 2008; and in revised form 13 Oct. 2008; accepted 2 Dec. 2008.
- forensic science;
- DNA typing;
- DNA degradation;
- ligase detection reaction;
- polymerase chain reaction
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 <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.