Résumé
OBJECTIVE@#To establish a simple, fast and economical technique for multiplex-typing SNPs and to explore its potential forensic application.@*METHODS@#Five Y-SNP loci (IMS-JST164520, IMS-JST021354, IMS-JST003305, M119 and M134) were selected and the allele specific primers of each locus were designed with the universal reporter primers tailed at their 5' end. Alleles of these loci were amplified first by allele specific primers, then amplified by universal reporter primers tagged by fluorescent dye.@*RESULTS@#A fluorescent-multiplex PCR system of the five Y-SNP loci was established. The typing results showed that two different colors of product peaks denoted two different alleles of a SNP locus, and the fragment sizes of alleles among different SNP loci were different. The haplotype diversity of these five loci was estimated to be 0.8655 in Wuhan Han population.@*CONCLUSION@#The multiplex-typing SNPs based on the universal reporter primers is a simple, fast, and economical technique, and may have good application value in forensic medicine.
Sujets)
Femelle , Humains , Mâle , Allèles , Asiatiques/génétique , Chine/ethnologie , Chromosomes Y humains/génétique , Amorces ADN , Génétique légale , Fréquence d'allèle , Marqueurs génétiques , Génétique des populations , Haplotypes , Techniques d'amplification d'acides nucléiques/méthodes , Polymorphisme de nucléotide simple/génétiqueRésumé
OBJECTIVE@#To evaluate the validity of Differential Extraction Kit in isolating spermatozoa and epithelial cell DNA from mixture samples.@*METHODS@#Selective lysis of spermatid and epithelial cells combined with paramagnetic particle method were applied to extract the DNA from the mock samples under controlled conditions and forensic case samples, and template DNA were analyzed by STR genotype method.@*RESULTS@#This Differential Extraction Kit is efficient to obtain high quality spermatid and epithelial cell DNA from the mixture samples with different proportion of sperm to epithelial cell.@*CONCLUSION@#The Differential Extraction Kit can be applied in DNA extraction for mixed stain from forensic sexual assault samples.
Sujets)
Femelle , Humains , Mâle , Séparation cellulaire/méthodes , ADN/isolement et purification , Cellules épithéliales/cytologie , Médecine légale , Réaction de polymérisation en chaîne/méthodes , Trousses de réactifs pour diagnostic , Sperme/cytologie , Infractions sexuelles , Spermatozoïdes/cytologieRésumé
It's always a challenge to type from highly degraded biological remains. PCR-based STR genotyping is helpful and valuable for such degraded samples like bones, teeth et al, but the typing results are sometimes unstable or wrong. Here the methods for solving the problems and improving reproducibility are reviewed.