To establish the rapid PCR amplification program and system and to verify the technical indexes.PCR multiplex and capillary electrophoresis detection of 24 autosomal STR loci and one Y-STR loci using the 6-color fluorescence marking technology, as well as A melogenin and Y-InDel. Meanwhile, sensitivity, specificity, identity, stability, mixing and a batch of sample tests were investigated, and the genotype of various routine samples and degraded, exfoliated cell samples were observed.The sensitivity of the system was 0.062 5 ng. In addition, the genotype could be detected accurately only around 65 min via rapid amplification. The species-specificity was high and the genotyping of all kinds of dry blood specimens of filter paper and mixed, degraded, exfoliated cell samples were accurate.The rapid amplification system can significantly improve the detection rate, and obtain accurate and stable genotyping results, which may be important implications for the establishment of STR database and study on population genetics and forensic identification.
Abstract A total of 101 blood samples were collected from unrelated males of Tibetan ethnic group in Lasa of China. DNA was extracted using Chelex method (1). The allelic variation at the three Y-STR loci named as DYS434, DYS443, and DYS456, were analyzed by PCR amplication with multiplex system. Multiplex PCR amplifications were performed in a 37.5 μL containing 2–4 ng human genome DNA, 7.5 μL dNTP (1 mmol/mL), 3μ, Taq polymerase, 3.75 μL 10 × buffer (Mg2+ 1.5 mmol/l), 3.75 μL BSA (1.6 μg/mL), 0.3 μL each primers (50 nmol/mL).
OBJECTIVE To understand the allele structure and genetic polymorphism at three STR loci on chromosome X in Chinese Han population and make an evalution of their forensic application. METHODS EDTA-blood samples were collected from the unrelated individuals in Chengdu, China. After being extracted with Chelex method, the DNA samples were amplified by PCR technique. The PCR products were analyzed by PAG electrophoresis and the approach of automated fluorescence detection. Hardy-Weinberg equilibrium of females was tested and every forensic interested value was calculated. RESULTS The polymorphisms of all 3 STR loci were obtained from 100 unrelated females and 120 unrelated males from Chinese Han ethnic group. Chi-square tests on the genotype frequencies in females did not reveal deviations from Hardy-Weinberg equilibrium. CONCLUSION The obtained data are beneficial to understanding the population genetics of the three STR loci in Chinese Han population. For forensic genetics, the obtained data can be used to calculate the probabilities dealing with the paternity test and the individual identification.
To investigate the genetic polymorphisms of 18 autosomal short tandem repeats （STR） loci in Changsha Han population, and explore the population genetic relationships and evaluate its application value in forensic medicine.The DNA of 2 004 unrelated individuals in Changsha Han population were amplified using Goldeneye®DNA ID System BASIC, and the PCR products were analyzed by electrophoresis using 3130xl genetic analyzer. The fragment sizes of alleles were analyzed subsequently by GeneMapper®ID v3.2. The frequency data and forensic genetic parameters ［observed heterozygosity （Ho）, expected heterozygosity （He）, power of discrimination （DP） and polymorphic information content （PIC）］ of 18 STR loci were statistically analyzed. Total probability of discrimination （TDP）, probability of exclusion in trio cases （PEtrio） and probability of exclusion in duo cases （PEduo） were calculated by Cervus 3.0. Hardy-Weinberg equilibrium and linkage disequilibrium of the loci were detected by Arlequin v3.5. The results were compared with the available data of other populations from different races and regions.The power of discrimination （DP）, and the polymorphic information content （PIC） of each locus of Changsha Han population ranged from 0.783 6 to 0.987 9 and 0.549 4 to 0.914 5, respectively. The TDP, cumulative probability of exclusion in trio cases （CPEtrio） and cumulative probability of exclusion in duo cases （CPEduo） were 0.999 999 999 999 999 999 999 865 2, 0.999 999 979 and 0.999 988 325, respectively. According to the Nei's DA genetic distance, the genetic distance between Changsha Han and Hunan Han populations was the smallest （0.014 1）, while it was the largest （0.041 8） between Changsha Han and Xinjiang Kazakh populations.The 18 STR loci shows abundant genetic polymorphisms in Changsha Han population. The study of genetic diversity among different populations has an important meaning for the research of their origins, migrations and their relationships.长沙汉族18个常染色体STR基因座的遗传多态性.调查长沙汉族群体18个常染色体短串联重复（short tandem repeat，STR）序列遗传多态性，探讨其群体遗传学关系及法医学应用价值。.应用Goldeneye®DNA身份鉴定系统BASIC，对长沙地区2 004名汉族无关个体血样DNA进行扩增，3130xl基因分析仪电泳分析，GeneMapper®ID v3.2软件分析等位基因片段大小。统计分析18个STR基因座的频率数据和群体遗传学参数（观察杂合度、期望杂合度、个体识别率、多态信息含量），采用Cervus 3.0计算累积个体识别率、三联体非父排除率和二联体非父排除率，采用Arlequin v3.5进行各基因座Hardy-Weinberg平衡及连锁不平衡检验，并与其他地区已有人群数据进行比较。.长沙汉族各基因座个体识别率为0.783 6~0.987 9，多态信息含量为0.549 4~0.914 5。累积个体识别率、三联体非父排除率和二联体非父排除率分别为0.999 999 999 999 999 999 999 865 2、0.999 999 979和0.999 988 325。根据Nei的DA遗传距离发现，长沙汉族与湖南汉族遗传距离最近（0.014 1），与新疆哈萨克族的遗传距离相对最远（0.041 8）。.这18个STR基因座在长沙汉族群体中具有丰富的遗传多态性。研究不同民族群体的遗传多态性对了解他们的起源、迁移以及相互关系有重要的意义。.法医遗传学；多态现象，遗传；短串联重复；常染色体；遗传距离；汉族；长沙.
To study genetic polymorphisms of 20 Y-chromosomal short tandem repeats (STR) in Chaoshan Han population, and to evaluate their value in forensic science.Twenty Y-specific STR loci (DYS434, Y-GATA-A10, Y-GATA-H4, DYS438, DYS439, DYS443, DYS444, DYS446, DYS447, DYS448, DYS456, DYS458, DYS460, DYS520, DYS531, DYS557, DYS622, DYS630, DYS635 and DYS709) were amplified by using three fluorescence-labeled multiplex PCR systems and were analyzed by ABI310 genetic analyzer. One hundred and fifty-eight unrelated male individuals of Han population in Chaoshan area were investigated to determine the distribution of allele frequencies and haplotype.The Y-STR multiplexes developed had followed the published nomenclature and ISFG guidelines for STR analysis. Gene diversity ranged from 0.2506 at DYS434 to 0.8034 at DYS447. A total of 157 different haplotypes were observed, and among these, 156 were unique, while 1 was found for two times. The haplotype diversity value calculated from all 20 loci combined was 0.999998. None of Y-STR allele mutation was observed in the 30 father/ son pairs confirmed by autosomal STR analysis.The results indicate that the 20 Y-STR loci are highly polymorphic and fathership genealogy inheritance are stable. The three fluorescence-labeled multiplex amplification systems that we constructed are suitable for forensic individual identification and paternity testing in Chaoshan area.
We analysed the forensic characteristics and substructure of the Handan Han population based on 36 Y-STR (short tandem repeat) and Y-SNP (single nucleotide polymorphism) markers. The two most dominant haplogroups in Handan Han, O2a2b1a1a1-F8 (17.95%) and O2a2b1a2a1a (21.51%), and their abundant downstream branches, reflected the strong expansion of the precursor of the Hans in Handan. The present results enrich the forensic database and explore the genetic relationships between Handan Han and other neighbouring and/or linguistically close populations, which suggests that the current concise overview of the Han intricate substructure remains oversimplified.
Abstract Blood samples were collected from unrelated individuals of Chinese Han ethnic group in Chengdu of China. DNA was extracted using Chelex method (1). PCR amplification conditions can be accessed at http://www.legalmed.org/dna/DXS6797.htm. The volume of PCR reaction for each locus was 37.5 µL. The PCR products were analyzed by horizontal non-denaturing polyacrylamide gel electrophoresis with discontinuous buffer system and visualized by silver staining (2). Data of female population genetics and forensic science were analyzed using POWERSTATS program (3). The complete dataset can be accessed at http://www. legalmed.org/dna/DXS6797.htm.
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