RESUMO
Los microsatélites o STR por su sigla en inglés (Short Tandem Repeat) son repeticiones de secuencias cortas de nucleótidos a través del DNA, de gran importancia debido a su carácter polimórfico que permite su utilización en la identificación genética de individuos o poblacionales. En este trabajo se investigaron los perfiles genéticos de una muestra de poblaciones humanas del altiplano cundiboyacense en Colombia; se usaron los marcadores STR's D2S1338, D19S433, PENTA D, PENTA E y SE-33. Éstos cinco STR's fueron amplificados mediante reacción en cadena de la polimerasa (PCR), corridos con electroforesis capilar, y tipificados con los programas Genscan y Genotyper. Como resultado se reportan las frecuencias alélicas de los cinco microsatélites (no reportados anteriormente para esta región). Se encontró que los loci D19S433 y SE-33 no están en equilibrio de Hardy-Weinberg; y que el Penta E tiene el mayor poder de discriminación. La distribución de las frecuencias alélicas para los cinco marcadores, en las dos poblaciones analizadas mostró que no existen diferencias estadísticas significativas entre ellas.
Microsatellites (STR's) are short tandem repeat sequences of nucleotides through DNA, very important for their polymorphic character that allow their utilization in population genetic identification. The STR's markers D2S1338, D19S433, PENTA D, PENTA E y SE33 were used to do the genetic profiles of human populations from the Cundiboyacense region in Colombia. The five STR's were amplified by PCR (polymerase chain reaction), indentified with capillary electrophoresis, and typified with Genscan and Genotyper software. Allelic frequencies of the five microsatellites are reported (not previously reported for this region). It was found that loci D19S433 y SE-33 are not in Hardy-Weinberg equilibrium; and Penta E provides the greatest power of discrimination. The allelic frequencies distribution to these five markers, in the two populations studied show that it does not exist statistically significant differences between them.
RESUMO
A duplex PCR reactions were designed for two pentameric (Penta D, Penta E) short tandem repeats (STR) loci to study the allele frequencies and the applicability of genetic variation in these loci to forensic case works. For 323 unrelated Koreans DNA was extracted using Qiagen DNA blood kit. The fluorescently labeled amplified products were analyzed by 8M urea, 5% long range gel electrophoresis in ABI-377 sequencer and followed data analysis by Genescan Analysis and Genotyper DNA fragment analysis software. The PD, PIC and PE were above 0.92, 0.76 and 0.66, respectively. No deviation from Hardy-Weinberg equilibrium was observed of Penta D locus (p=0.35, observed number>1).
Assuntos
DNA , Eletroforese , Frequência do Gene , Variação Genética , Repetições de Microssatélites , Reação em Cadeia da Polimerase , Estatística como Assunto , UreiaRESUMO
Objective To study the primer design for Penta D and Penta E genotyping and investigate the genetic polymorphisms of these two loci in Chinese Han population in Wuhan.Methods 281 unrelated Chinese individuals living in Wuhan were typed by hot-start PCR with re-designed Penta D and Penta E genotyping primers and PAGE technique,and the results were compared with those by the PowerPlex TM 16 system of Promega.Results The amplified fragment size of Penta D and Penta E loci with re-designed primers ranged between 153～198bp and 107～212bp respectively,which were consistent with the results obtained by the re-designed primers and PowerPlex TM 16 system,and the re-designed primers had a higher sensitivity than PowerPlex TM 16 system (0.2ng vs 0.5ng) by silver staining.10 and 21 alleles were observed for Penta D and Penta E in Chinese Han population in Wuhan,and the genotype distributions of the two loci were in accordance with Hardy-Weinberg equilibrium.Family studies confirmed Mendelian inheritance of alleles.The power of discrimination (DP) for Penta D and Penta E were 0.926 2 and 0.986 0,and the power of exclusion (PE) were 0.665 1 and 0.832 5 respectively.Conclusion The re-designed primers for Penta D and Penta E genotyping are reliable.These two loci are highly polymorphic in Chinese Han population and can be used in forensic identification and paternity test.