RESUMEN
The development of massively parallel sequencing (MPS) has quickly changed forensic short tandem repeat (STR) genotyping. By providing detailed sequence information, MPS technology may be used as an alternative or additional method to overcome the limitations of capillary electrophoresis-based STR profiling. Most current NGS processes are labour-intensive with regard to library preparation and require high-quality DNA template. In this study, a 16-plex STR typing system (SeqType®R16) was used to achieve direct library preparation without DNA extraction and adaptor ligation. The efficiency of this system was tested in 601 individuals, including 593 old blood samples from the Chinese Han population and eight positive controls. It took approximately 4 hours for library preparation, including blood direct multiplex PCR (1.5 hours), mixing of the product (15 minutes), single tube purification (2 hours) and quantification (15 minutes). The results showed that MPS presented a broader allele range and higher discrimination power. Except for FGA and D19S433, the allele number almost doubled or more than doubled at all complex STR loci and simple STR loci, including D13S317, D16S539, D5S818, and D7S820. The range of discrimination power increased from 0.8008-0.9572 to 0.8401-0.9753, and the culminated matching probability decreased from 1.7 × 10-15 to 1.1 × 10-17.
Asunto(s)
Pueblo Asiatico/genética , Manchas de Sangre , Genética Forense , Repeticiones de Microsatélite , Polimorfismo Genético , Femenino , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , MasculinoRESUMEN
OBJECTIVE: In order to provide a theoretical foundation for the medically effective ingredient-based selection of elite cultivars in Comnus officinalis, a study has been conducted on the variation in medicinal effective ingredients of the fruit from the genuine producing areas, the correlation among effective medicinal ingredients and the correlation between effective ingredients and fruit shape as well as nutritional indexes. METHOD: The completely mature fruit was collected from the genuine producing areas Chunan county and Lin'an city of Zhejiang province. The contents of colchicine, ursolic acid and oleanolic acid were determined by HPLC, and vertical diameter/transversal diameter of the fruits, soluble solid matter and percentage of fresh flesh to the fruit were also measured. RESULT: (1) Ursolic acid, oleanolic acid, and colchicine in fruits ranged from 0.1010% to 0.4786%, 0.0149% to 0.1274% and 0.59% to 2.30%, respectively, and their RSD were 34.33%, 40.48% and 28.50%, respectively. (2) The correlation between effective ingredients and that between effective ingredients and fruit shape as well as nutritional indexes were as follows: the content of ursolic acid was significantly correlated with that of oleanolic acid with a correlation coefficient of 0.9796; both ursolic acid and oleanolic acid were in significantly negative correlation with soluble solid matter with a correlation coefficient of -0.5544 and -0.5118, respectively; colchicine was significantly associated with soluble solid matter with a correlation coefficient of 0.2412; colchicine, ursolic acid and oleanolic acid were in significantly negative correlation with the percentage of fresh flesh with a correlation coefficient of -0.2507, -0.2443 and -0.2406, respectively; three effective ingredients showed no correlation with the ratio of vertical diameter to transversal diameter of the fruit. CONCLUSION: There is a significant difference in effective ingredients among individual trees, which means that there is a big potential for selection of cultivars. Individual tree-based selection should be mainly adopted when effective ingredients are used as a main index in selection on the basis of the correlation among effective ingredients and that between effective ingredients and fruit shape as well as nutritional indexes, while ursolic acid could be combined with oleanolic acid to be used as an index and a preliminary screen could be conducted using soluble solid matter.