Validation of the DNATyper™15 PCR Genotyping System for Forensic Application
Jian Ye,Chengtao Jiang,Xingchun Zhao,Le Wang,Cai-Xia Li,Anquan Ji,Li Yuan,Jing Sun,Shuaifeng Chen +8 more
- 01 Jan 2015
- Vol. 1, Iss: 1, pp 8
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TL;DR: The performance of the DNATyper™15 system on casework samples was compared with that of two widely used STR amplification kits, Identifiler™ and PowerPlex 16 ®, and the performances were comparable in terms of sensitivity and the ability to genotype the mixed samples and case-type samples.
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Abstract: We describe the optimization and validation of the DNATyper™15 multiplex polymerase chain reaction (PCR) genotyping system for autosomal short tandem repeat (STR) amplification at 14 autosomal loci (D6S1043, D21S11, D7S820, CSF1PO, D2S1338, D3S1358, D13S317, D8S1179, D16S539, Penta E, D5S818, vWA, D18S51, and FGA) and amelogenin, a sex-determining locus. Several DNATyper™15 assay variables were optimized, including hot start Taq polymerase concentration, Taq polymerase activation time, magnesium concentration, primer concentration, annealing temperature, reaction volume, and cycle number. The performance of the assay was validated with respect to species specificity, sensitivity to template concentration, stability, accuracy, influence of the DNA extraction methods, and the ability to genotype the mixture samples. The performance of the DNATyper™15 system on casework samples was compared with that of two widely used STR amplification kits, Identifiler™ (Applied Biosystems, Carlsbad, CA, USA) and PowerPlex 16 ® (Promega, Madison, WI, USA). The conditions for PCR-based DNATyper™15 genotyping were optimized. Contamination from forensically relevant nonhuman DNA was not found to impact genotyping results, and full profiles were generated for all the reactions containing ≥ 0.125 ng of DNA template. No significant difference in performance was observed even after the DNATyper™15 assay components were subjected to 20 freeze-thaw cycles. The performances of DNATyper™15, Identifiler™, and PowerPlex 16 ® were comparable in terms of sensitivity and the ability to genotype the mixed samples and case-type samples, with the assays giving the same genotyping results for all the shared loci. The DNA extraction methods did not affect the performance of any of the systems. Our results demonstrate that the DNATyper™15 system is suitable for genotyping in both forensic DNA database work and case-type samples.
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Citations
Genetic Diversity and Phylogenetic Differentiation of Southwestern Chinese Han: a comprehensive and comparative analysis on 21 non-CODIS STRs
TL;DR: Analysis of molecular variance (AMOVA), principal component analysis (PCA), multidimensional scaling plots (MDS) and phylogenetic analysis consistently demonstrated that the Southwest Han population had a close genetic relationship with the geographically close population (Hunan Han) and kept a distant Genetic relationship with some ethnic groups.
Validation study of a 15-plex rapid STR amplification system for human identification
TL;DR: It is found that the rapid 15-plex PCR system could shorten the amplification time to 37min and proved that it provides an alternative method for conventional PCR in human identification detection.
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PCR-RFLP identification of meat from red deer, sika deer, roe deer, fallow deer, mouflon, wild boar, hare and cattle.
Michaela Nesvadbova,Michaela Nesvadbova,Gabriela Bořilová,Gabriela Bořilová,Radka Hulánková,Radka Hulánková +5 more
TL;DR: New polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) based on digestion of PCR products with two restriction enzymes, MboII and AciI, have been developed for the specific identification of raw and heat-processed meat from red deer, sika deer, and cattle.