scholarly journals Analysis of microsatellite markers D18S70 and d20S116 in DNA isolated from dentin: Use in forensic medicine

2009 ◽  
Vol 137 (1-2) ◽  
pp. 43-46 ◽  
Author(s):  
Dragana Puzovic ◽  
Branka Popovic ◽  
Ivana Novakovic ◽  
Jelena Milasin

Introduction. Short tandem repeats and more specifically microsatellites represent a powerful tool in forensic medicine. In the past years, they have been extensively used in human identification and paternity testing. Objective The aim of the present study was to analyze two microsatellite markers in the Serbian population, i.e. to determine the number of alleles and the relevant forensic parameters. Methods. DNA was isolated from teeth samples using standard proteinase K digestion and phenol/chloroform alcohol extraction. PCR products were analyzed on polyacrilamide gels and visualized by AgNO3 staining. Forensic parameters were calculated using the Cervus software. Results. The loci D18S70 and D20S116 were analyzed on a sample of 70 unrelated, healthy adult individuals from Serbia. The number of alleles was determined and Hardy Weinberg equilibrium was confirmed for both loci. D18S70 and D20S116 demonstrated 6 and 8 alleles, respectively. The power of discrimination (PD) and the power of exclusion (PE) for the tested STR loci, D18S70 and D20S116 were 0.92 (PD), 0.41 (PE) and 0.95 (PD), 0.480 (PE), respectively. Conclusion. According to the presented data, D18S70 and D20S116 are most informative markers. Based on allelic frequencies and statistical parameters for forensic testing, it may be suggested that these two microsatellites represent useful markers for individual identification and parentage analysis in the Serbian population.

PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e5946 ◽  
Author(s):  
Jie Pei ◽  
Pengjia Bao ◽  
Min Chu ◽  
Chunnian Liang ◽  
Xuezhi Ding ◽  
...  

Background Yak (Bos grunniens) is the most important domestic animal for people living at high altitudes. Yak ordinarily feed by grazing, and this behavior impacts the accuracy of the pedigree record because it is difficult to control mating in grazing yak. This study aimed to evaluate the pedigree system and individual identification in polled yak. Methods A total of 71 microsatellite loci were selected from the literature, mostly from the studies on cattle. A total of 35 microsatellite loci generated excellent PCR results and were evaluated for the parentage testing and individual identification of 236 unrelated polled yaks. A total of 17 of these 35 microsatellite loci had polymorphic information content (PIC) values greater than 0.5, and these loci were in Hardy–Weinberg equilibrium without linkage disequilibrium. Results Using multiplex PCR, capillary electrophoresis, and genotyping, very high exclusion probabilities were obtained for the combined core set of 17 loci. The exclusion probability (PE) for one candidate parent when the genotype of the other parent is not known was 0.99718116. PE for one candidate parent when the genotype of the other parent is known was 0.99997381. PE for a known candidate parent pair was 0.99999998. The combined PEI (PE for identity of two unrelated individuals) and PESI (PE for identity of two siblings) were >0.99999999 and 0.99999899, respectively. These findings indicated that the combination of 17 microsatellite markers could be useful for efficient and reliable parentage testing and individual identification in polled yak. Discussion Many microsatellite loci have been investigated for cattle paternity testing. Nevertheless, these loci cannot be directly applied to yak identification because the two bovid species have different genomic sequences and organization. A total of 17 loci were selected from 71 microsatellite loci based on efficient amplification, unambiguous genotyping, and high PIC values for polled yaks, and were suitable for parentage analysis in polled yak populations.


2009 ◽  
Vol 6 (5) ◽  
Author(s):  
Cristina Pinto

AbstractThrough the last decade there was an enormous revolution in the field of forensic genetic.The Author reviews some of the methodologies used in the definitions of DNA profiling tackling the principles of recombinant DNA techniques. The potentiality of polymorphic DNA fragments in vertebrates is focused as well as the revolution implied in forensic medicine. The resource to DNA-DNA hybridization combined to oligonucleotide probes is emphasized leading to the production of an individual bar code with the resource of genomic polymorphism which leads to a pattern known as genetic fingerprinting. Other techniques for individual identification and paternity testing are focused as well as the use of short tandem repeats (STR's). Mitochondrial DNA sequencing use to complement nuclear DNA typing may also be profitable in certain instances. Relevant problems within the context of the use of these techniques in forensic medicine and law suits are discussed. Final considerations viewing the resource to DNA technology within the scope of the last two decades are referred regarding the resource to DNA profiles not only in the US but in Europe in general and in Portugal in special having lead to compensation and uncover of justice errors.


2020 ◽  
pp. 002580242096500
Author(s):  
Supakit Khacha-ananda ◽  
Phatcharin Mahawong

Short tandem repeats (STRs) are widely used as DNA markers in paternity testing and criminal investigations because of their high genetic polymorphism among individuals in population. However, many factors influence genetic variations of STRs. Therefore, understanding STR information within individual populations could provide database and scientifically reliable STR genotyping for forensic genetic purposes. We aimed to examine allele frequencies of X-STRs, including some forensic parameters, in a northern Thai population. A retrospective descriptive study was conducted by collecting X-STR data from unrelated individuals living in a northern region of Thailand. The allele frequency and forensic parameters – for example polymorphism information content (PIC), power of discrimination in females and males (PDf and PDm), mean exclusion chance (MEC) and haplotype frequency – were calculated. The Hardy–Weinberg equilibrium was analysed. A total of 132 alleles were observed, with corresponding allele frequency ranging from 0.0064 to 0.4904. The PIC of all loci was >0.6, representing high genetic polymorphism, except DXS8378 and DXS7423. Notably, DXS10135 was the most diverse loci with the highest PD and MEC, while DXS7423 was the least polymorphic marker with the lowest PD and MEC. The highest haplotype diversity in male data was on linkage group III (DXS10101-DXS10103-HPRTB) by 0.9895. The genetic distance analysis demonstrated that the northern Thai population had a close relationship with Taiwanese (DA = 0.023). There are no significant deviations among the Hardy–Weinberg equilibrium except DXS10148. This study has established a northern Thai X-STRs reference database to be used as a tool for forensic genetic purposes.


Genes ◽  
2020 ◽  
Vol 11 (7) ◽  
pp. 782
Author(s):  
Eva Bazsalovicsová ◽  
Gabriel Minárik ◽  
Katarína Šoltys ◽  
Alžbeta Radačovská ◽  
Jesper A. Kuhn ◽  
...  

Dibothriocephalus dendriticus is one of the causative agents of the fish-borne zoonosis diphyllobothriosis. Polymorphic microsatellite markers were originally developed for future genetic studies using microsatellite library screening and next-generation sequencing (NGS). Out of 128 microsatellite candidates selected after NGS analysis, 126 yielded PCR products of the expected size. A declared repetitive motif was confirmed in 92 loci by Sanger sequencing. The level of polymorphism was tested by fragment analysis. Statistical tests for observed and expected heterozygosities and deviations from Hardy–Weinberg equilibrium revealed 14 polymorphic microsatellite loci suitable for studies on the finer genetic structure of global populations of D. dendriticus.


Genes ◽  
2021 ◽  
Vol 12 (7) ◽  
pp. 1042
Author(s):  
Zhuoying Weng ◽  
Yang Yang ◽  
Xi Wang ◽  
Lina Wu ◽  
Sijie Hua ◽  
...  

Pedigree information is necessary for the maintenance of diversity for wild and captive populations. Accurate pedigree is determined by molecular marker-based parentage analysis, which may be influenced by the polymorphism and number of markers, integrity of samples, relatedness of parents, or different analysis programs. Here, we described the first development of 208 single nucleotide polymorphisms (SNPs) and 11 microsatellites for giant grouper (Epinephelus lanceolatus) taking advantage of Genotyping-by-sequencing (GBS), and compared the power of SNPs and microsatellites for parentage and relatedness analysis, based on a mixed family composed of 4 candidate females, 4 candidate males and 289 offspring. CERVUS, PAPA and COLONY were used for mutually verification. We found that SNPs had a better potential for relatedness estimation, exclusion of non-parentage and individual identification than microsatellites, and > 98% accuracy of parentage assignment could be achieved by 100 polymorphic SNPs (MAF cut-off < 0.4) or 10 polymorphic microsatellites (mean Ho = 0.821, mean PIC = 0.651). This study provides a reference for the development of molecular markers for parentage analysis taking advantage of next-generation sequencing, and contributes to the molecular breeding, fishery management and population conservation.


Author(s):  
Purabi Kaushik ◽  
Jnyanashree Saikia ◽  
Kabitabala Kalita ◽  
Rajjyoti Deka ◽  
J. Saharia

Background: Duck farming plays a significant role, next to chicken in the socio-economic uplift men of the rural farmers of North-East India. Pati duck is the most common duck breed in the Brahmaputra valley of Assam and the other common variety reared in North-East India is Chara-Chambeli, however it originatein Kerala. Genetic characterization plays a significant role for formulation of breeding strategies for improvement of any breed. Microsatellites are codominant in nature and are highly polymorphic. High level of allelic variation, co-dominant mode of inheritance and potential for automated analysis make them an excellent tool for genotyping, mapping and genetic characterization. Pati duck is the most common duck breed in the Brahmaputra valley of Assam and the other common variety reared in North-East India is Chara-Chambeli however its origin in Kerela. Therefore, the present investigation was carried out to characterize these two duck breeds using microsatellite markers to evaluate the genetic diversity in these two duck population. Methods: For the present study, Blood sample were collected from 50 Patiducks and 50 Chara-Chambeli ducks from different parts of North-East India. Assessment of genetic characterization of duck breeds of north-east region were carried out using 16 microsatellite markers and population genetics analysis were done by POPGENE software. Result: In the present study, all the studied loci were highly polymorphic. Analysis generated a total of 41 microsatellite alleles. The number of observed alleles (Na) with an overall mean of 1.93±0.258. However, the effective number of alleles (Ne) with a mean of 1.6933±0.2712. The Shannon’s information index was found to a mean value of 0.5685±0.1693. The overall means for observed (HO) and expected (He) heterozygosities were 0.2889±0.2477 and 0.5289±0.0853, respectively. The chi-square (χ2) test for Hardy-Weinberg equilibrium revealed that all the loci are in within Hardy-Weinberg equilibrium.


PeerJ ◽  
2016 ◽  
Vol 4 ◽  
pp. e2416 ◽  
Author(s):  
Catherine Jan ◽  
Luca Fumagalli

The parrot family represents one of the bird group with the largest number of endangered species, as a result of habitat destruction and illegal trade. This illicit traffic involves the smuggling of eggs and animals, and the laundering through captive breeding facilities of wild-caught animals. Despite the huge potential of wildlife DNA forensics to determine with conclusive evidence illegal trade, current usage of DNA profiling approaches in parrots has been limited by the lack of suitable molecular markers specifically developed for the focal species and by low cross-species polymorphism. In this study, we isolated DNA microsatellite markers in seven parrot species threatened with extinction (Amazona brasiliensis,A. oratrix,A. pretrei,A. rhodocorytha,Anodorhynchus leari,Ara rubrogenysandPrimolius couloni). From an enriched genomic library followed by 454 pyrosequencing, we characterized a total of 106 polymorphic microsatellite markers (mostly tetranucleotides) in the seven species and tested them across an average number of 19 individuals per species. The mean number of alleles per species and across loci varied from 6.4 to 8.3, with the mean observed heterozygosities ranging from 0.65 to 0.84. Identity and parentage exclusion probabilities were highly discriminatory. The high variability displayed by these microsatellite loci demonstrates their potential utility to perform individual genotyping and parentage analyses, in order to develop a DNA testing framework to determine illegal traffic in these threatened species.


2012 ◽  
Vol 10 (10) ◽  
pp. 20-23 ◽  
Author(s):  
Dinesh Kumar Jha ◽  
Luis Javier Martinez Gonzalez ◽  
Jiwan Prasad Rijal ◽  
Bhinu Shova Tuladhar ◽  
Nirajan Thapa Chhetri

Allele frequencies for the 15 STR loci in the AmpFlSTR® Identifiler® and statistical parameters were estimated from a sample of 233 unrelated individuals from different area of Nepal. A total of 161 alleles were found, with corresponding allelic frequencies ranging from 0.0021 to 0.4077. The MP, PD, PIC, PE, TPI, Ho and He ranged from 0.0282 to 0.1511, 0.8489 to 0.9717, 0.6478 to 0.8700, 0.4546 to 0.7631, 1.7651 to 4.3148, 0.7167 to 0.8841 and 0.7009 to 0.8833 respectively. Deviations from the Hardy–Weinberg Equilibrium were observed for D16S539, D18S51, D21S11 and TPOX Markers. One new allele has been detected in the process. Scientific World, Vol. 10, No. 10, July 2012 p20-23 DOI: http://dx.doi.org/10.3126/sw.v10i10.6856


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