![]() Overall, these thoroughly vetted full mtGenome population reference data can serve as a standard against which the quality and features of future mtGenome datasets (especially those developed via massively parallel sequencing) may be evaluated, and will provide a solid foundation for the generation of complete mtGenome haplotype frequency estimates for forensic applications. ![]() The ratio of nonsynonymous to synonymous changes among point heteroplasmies in the protein-coding genes (1:1.3) and average pathogenicity scores in comparison to data reported for complete substitutions in previous studies seem to provide some additional support for the role of purifying selection in the evolution of the human mtGenome. Name : Description: DNA Sequence Assembler : DNA Baser is a tool for DNA Sequence Assembler, DNA sequence analysis, contig editing, and mutation detection.It also offers a powerful chromatogram viewer/editor. In the coding region, nearly 90% of individuals exhibited length heteroplasmy in the 12418-12425 adenine homopolymer and despite a relatively high rate of point heteroplasmy (23.8% of individuals across the entire molecule), coding region point heteroplasmies shared by more than one individual were notably absent, and transversion-type heteroplasmies were extremely rare. Inferred maternal biogeographic ancestry frequencies for each population and heteroplasmy rates in the control region were generally consistent with published datasets. For all three populations, nearly complete resolution of the haplotypes was achieved with full mtGenome sequences: 90.3–98.8% of haplotypes were unique per population, an improvement of 7.7–29.2% over control region sequencing alone, and zero haplotypes overlapped between populations. Data review followed an intensive multi-step strategy that included a minimum of three independent reviews of the raw data at two laboratories repeat screenings of all insertions, deletions, heteroplasmies, transversions and any additional private mutations and a check for phylogenetic feasibility. We finally assembled all Sanger sequencing and NGS sequence data using Sequencher 5.1 (Gene Code, Ann Arbor, MI, USA) and determined the complete mtDNA sequence of Py. Data production utilized an 8-amplicon, 135 sequencing reaction Sanger-based protocol, performed in semi-automated fashion on robotic instrumentation. Almost the complete mtDNA sequence was produced by clone sequencing, gap-filling PCR, and primer walking and was compared with contig sequences from the NGS system. population groups (African American, Caucasian and Hispanic) from anonymized, randomly-sampled specimens. To address this deficiency, we have developed 588 complete mtGenome haplotypes, spanning three U.S. Though investigations into the use of massively parallel sequencing technologies for the generation of complete mitochondrial genome (mtGenome) profiles from difficult forensic specimens are well underway in multiple laboratories, the high quality population reference data necessary to support full mtGenome typing in the forensic context are lacking.
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