Resumen de A LDR-PCR approach for multiplex polymorphisms genotyping of severely degraded DNA with fragment sizes <100 bp
Zhen-hua Zhang, Bao-Jie Wang, Hong-Yu Guan, Hao Pang, Jin-Feng Xuan
Reducing amplicon sizes has become a major strategy for analyzing degraded DNA typical of forensic samples. However, ampli-con sizes in current mini-short tandem repeat-polymerase chain reaction (PCR) and mini-sequencing assays are still not suitable for analysis ofseverely degraded DNA. In this study, we present a multiplex typing method that couples ligase detection reaction with PCR that can be used toidentify single nucleotide polymorphisms and small-scale insertion ⁄ deletions in a sample of severely fragmented DNA. This method adopts thermo-stable ligation for allele discrimination and subsequent PCR for signal enhancement. In this study, four polymorphic loci were used to assess the abil-ity of this technique to discriminate alleles in an artificially degraded sample of DNA with fragment sizes <100 bp. Our results showed clear allelicdiscrimination of single or multiple loci, suggesting that this method might aid in the analysis of extremely degraded samples in which allelic dropout of larger fragments is observed
