RESUMEN
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.
Asunto(s)
Alelos , Cromosomas Humanos Y/genética , Cartilla de ADN , Técnicas de Amplificación de Ácido Nucleico/métodos , Polimorfismo de Nucleótido Simple/genética , Pueblo Asiatico/genética , China/etnología , Femenino , Genética Forense , Frecuencia de los Genes , Marcadores Genéticos , Genética de Población , Haplotipos , Humanos , MasculinoRESUMEN
To search polymorphic Y chromosome biallelic markers in Chinese Han population, and obtain their population genetic data. Genotyping of 23 biallelic markers on human Y chromosome (M7, M9, M50, M88, M89, M95, M111, M117, M119, M121, M122, M134, M159, M164, M175, M214, LINE1, MSY2, RPS4Y711, SRY465, IMS-JST164520, IMS-JST021354 and IMS-JST003305) were carried out in a sample of 160 unrelated Chinese male individuals living in Wuhan using fragment length discrepant allele specific PCR (FLDAS-PCR) and PAGE technique. In all 23 biallelic markers, genetic polymorphism were identified for 20 loci in Wuhan Han population except for M50, M159 and M164, and the ranges of gene diversity (GD) were 0.0126-0.4855. A total of 35 different haplogroups (Hg1-35) were observed and the haplogroup diversity (HD) was 0.9471. The haplogroups formed by 20 biallelic markers are highly polymorphic, and can be used in forensic science and population evolution studies.
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Pueblo Asiatico/genética , Cromosomas Humanos Y/genética , Marcadores Genéticos , Polimorfismo Genético , Alelos , Pueblo Asiatico/etnología , Haplotipos , Humanos , MasculinoRESUMEN
OBJECTIVE: To study the application of PCR-SSCP in forensic mtDNA typing. METHODS: Primers flanking the mtDNA HV-I and HV-II regions were designed. By PCR-SSCP techniques, 70 family trios and 140 unrelated Wuhan Han individuals were investigated and analyzed. RESULTS: In 70 family trios, the SSCP profiles in region HV-I and HV-II of children were not same to that of their fathers in 98.57% and 97.13% respectively but were identical with their mothers. In 140 unrelated Wuhan Han individuals, 21 haplotypes were found in HVI, GD = 0.9556; 16 haplotypes were found in HVII, GD = 0.9356. CONCLUSION: PCR-SSCP technique may be useful in forensic mtDNA typing, especially for screening the suspects.
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Dermatoglifia del ADN/métodos , ADN Mitocondrial/genética , Haplotipos , Reacción en Cadena de la Polimerasa/métodos , Polimorfismo Conformacional Retorcido-Simple , Cartilla de ADN , ADN Mitocondrial/sangre , Genética Forense/métodos , Humanos , Linaje , Análisis de Secuencia de ADNRESUMEN
OBJECTIVE: To establish a novel method for the multiplex analysis of the methylation and single nucleotide polymorphism (SNP). METHODS: The imprinted SNP rs220028 was chosen as a model. Genomic DNA, after being digested with methylation sensitive restriction enzyme, were typed by mutagenically separated PCR (MS-PCR). The polymorphism of restriction site was excluded by PCR-RFLP. RESULTS: By post-digestion MS-PCR, the methylated allele was detected selectively, the maternal origin of which was confirmed by pedigree analysis; A=0.5085, G=0.4915,PIC=0.3749. CONCLUSION: The multiplex analysis of methylation markers and SNP can be achieved by post-digestion MS-PCR. The imprinted SNP locus rs220028 is a potentially useful marker in screening Prader-Willi/Angelman syndrome.
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Marcadores Genéticos/genética , Reacción en Cadena de la Polimerasa/métodos , Polimorfismo de Nucleótido Simple , Metilación de ADN , Enzimas de Restricción del ADN/metabolismo , HumanosRESUMEN
DNA methylation is a post-replication modification that is predominantly found in cytosines of the dinucleotide sequence CpG. Epigenetic information is stored in the distribution of the modified base 5-methylcytosine. DNA methylation profiles represent a more chemically and biologically stable source of molecular diagnostic information than RNA or most proteins. Recent advances attest to the great promise of DNA methylation markers as powerful future tools in the clinic. In the past decade, DNA methylation analysis has been revolutionized by two technological advances--bisulphite modification of DNA and methylation-specific polymerase chain reaction (MSP). The methylation pattern of human genome is space-time specific, sex-specific, parent-of-origin specific and disease specific, providing us an alternative way to solve forensic problems.
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Metilación de ADN , Medicina Legal/métodos , Secuencia de Bases , Islas de CpG/genética , ADN/sangre , Dermatoglifia del ADN/métodos , Epigénesis Genética , Marcadores Genéticos , Genoma Humano , Humanos , Paternidad , Reacción en Cadena de la Polimerasa/métodosRESUMEN
OBJECTIVE: To establish a new method for single nucleotide polymorphism (SNP) typing based on allele specific PCR: fragment length discrepant allele specific PCR (FLDAS-PCR), and study the influence on specific extension by introducing a mismatch at the third or fourth 3'-terminal base of allele specific primers. METHODS: For SNP loci rs759117 and rs760887, two allele specific forward primers, with different length and a mismatch introduced at the third or fourth 3'-terminal base, and a public reverse primer were designed for SNP typing. The genotyping of SNP was determined by the two allele specific fragments different in size after polyacrylamide gel and silver staining. RESULTS: The different homozygote genotypes comprised a single band with different size respectively, and the heterozygote genotypes comprised two bands. Typing results were completely consistent with those by direct sequencing. Non-specific primer extension was decreased remarkably after introducing a mismatch at the third or fourth 3'-terminal base of allele specific primers, and the stringency of PCR reaction was cut down. CONCLUSION: FLDAS-PCR is a simple, rapid and efficient new method for SNP typing. During FLDAS-PCR, specific primers with a mismatch at the third or fourth 3'-terminal base have more power to identify two alleles.
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Alelos , ADN/genética , Polimorfismo de Nucleótido Simple , Disparidad de Par Base/genética , Cartilla de ADN , Electroforesis en Gel de Poliacrilamida , Genotipo , Humanos , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Sensibilidad y Especificidad , Análisis de Secuencia de ADN/métodosRESUMEN
This article review the application of chi-square test of various data handling methods and exact test in Hardy-Weinberg equilibrium testing of human genetic marker in population genetics. The importance of HWE-exact test in multiallelic system was emphasized, especially in the study of forensic VNTR and STR typing.
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Genética de Población/métodos , Modelos Genéticos , Alelos , Distribución de Chi-Cuadrado , Medicina Legal , Frecuencia de los Genes , Genotipo , Humanos , Funciones de Verosimilitud , Modelos EstadísticosRESUMEN
Nowadays, the injury in human mitochondrial DNA (mtDNA) is well known to accumulate in various tissues with age. It's significant to further investigate and then apply it to estimation of the age at parenchymas.
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Envejecimiento/fisiología , Daño del ADN/fisiología , ADN Mitocondrial/fisiología , Envejecimiento/genética , Disparidad de Par Base/genética , Fragmentación del ADN/genética , ADN Mitocondrial/genética , Eliminación de Gen , Humanos , Reacción en Cadena de la PolimerasaRESUMEN
OBJECTIVE: PGM1 genotyping by PCR-SSCP analysis. METHODS: Amplified genome DNA from 156 unrelated Han individuals living in Wuhan, PCR products for exon 4 and exon 8 of PGM1 were then analyzed by SSCP to detect the genotypes. RESULTS: 2 alleles and 3 genotypes were detected in exon 4 and 8 respectively. The discrimination power was 0.7318. PCR-SSCP analysis was suitable for determination of PGM1 genotypes from old blood and semen stains. CONCLUSION: PGM1 system typed by PCR-SSCP is useful for forensic identification.
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Pueblo Asiatico/genética , ADN/genética , Fosfoglucomutasa/genética , Reacción en Cadena de la Polimerasa/métodos , Polimorfismo Genético , Alelos , China , ADN/análisis , Frecuencia de los Genes , Genotipo , Humanos , Polimorfismo Conformacional Retorcido-Simple , Sensibilidad y EspecificidadRESUMEN
OBJECTIVE: The genetic polymorphism of two STR loci, D20S85 and D6S477, were studied in 280 unrelated Chinese individuals in Wuhan. METHODS: The PCR amplified products were analyzed by PAGE and silver staining. RESULTS: 10 and 9 alleles were observed in these two STR loci, and the discriminating power (DP) were 0.9085 and 0.9127 respectively. No deviations from Hardy-Weinberg equilibrium were found. The two STR loci had been successfully applied to individual identification and paternity testing. CONCLUSION: The results demonstrated that the two loci were useful for forensic identification.