scholarly journals Microsatellite analysis of cryopreserved stallion semen stored on FTA(R) paper : research communication

2002 ◽  
Vol 73 (4) ◽  
Author(s):  
M.L. Schulman ◽  
C.K. Harper ◽  
E. Bell ◽  
A. Nel ◽  
A.J. Guthrie
Keyword(s):  
Microsatellite Markers ◽  
Microsatellite Analysis ◽  
Proteinase K ◽  
Sperm Cells ◽  
Blood Samples ◽  
Research Communication ◽  
Full Profile ◽  
Dna Profiles ◽  
Semen Donor

The aim of this study was to establish and validate a method to permit microsatellite analysis of DNA profiles obtained from frozen-thawed stallion sperm cells. This would provide reliable and accurate verification of the identification of a semen donor. Ejaculates from 5 pony stallions were collected, processed and frozen in 0.5 m plastic straws. Aliquots of 100 m of the frozen-thawed semen thus obtained were either placed directly, or diluted (1 : 10 ; 1 : 100 ; and 1 : 1000) and placed on slides of FTA(R) paper. Similarly, blood samples obtained from each of the stallions were placed onto slides of FTA(R) paper. A punch was removed from each sample after drying. Each sample was mixed with FTA(R) purification reagent, Dithiothreitol and Proteinase K before incubation and processing. All samples were processed with a set of 13 microsatellite markers. Further analysis permitted a comparison of the DNA profiles of the frozen-thawed semen and the blood samples. A full profile of markers was obtained from the 1 : 10 and 1 : 100 dilutions of the frozen-thawed semen samples as well as from the blood samples. The DNA profiles from the frozen-thawed semen and blood samples obtained from the stallions matched in all cases.

scholarly journals Microsatellite Variability of Two Populations of Clarias gariepinus (Siluriformes, Clariidae) in Nigeria

2019 ◽  
Vol 53 (3) ◽  
pp. 195-208
Author(s):  
M. O. Awodiran ◽  
F. O. Adeniran ◽  
R. O. Akinwale ◽  
A. A. Akinwande
Keyword(s):  
Microsatellite Markers ◽  
Microsatellite Dna ◽  
Dna Analysis ◽  
Clarias Gariepinus ◽  
Microsatellite Analysis ◽  
Significant Differentiation ◽  
Different Populations ◽  
Genetic Signatures ◽  
Two Populations ◽  
River Niger

Abstract The study evaluated the genetic signatures of the fishes from the two populations and compared the pattern of differentiation of the two populations with a view to separating the species from the different populations into possible sub-species. Forty (40) specimens were collected from River Niger (Lokoja) and Asejire Resevoir. The DNA of the twenty (20) specimens from each population extracted from the muscle tissue using phenol-chloroform extraction (PCE) method was subjected to microsatellite DNA analysis. Seven (7) microsatellite markers (Cga01, Cga02, Cga03, Cga05, Cga06, Cga09 and Cga10) were used in the analysis. Microsatellite DNA analysis of the two populations revealed significant differentiation between the two populations as shown by the high values of heterozygosity, low level of inbreeding and non-conformance to Hardy-Weinberg’s equilibrium. It is concluded from the study that microsatellite analysis showed a high potentiality for separation of the populations.

scholarly journals Genetic relationship among indigenous sheep population of Bangladesh

2019 ◽  
Vol 48 (1) ◽  
pp. 17-22
Author(s):  
GK Deb ◽  
MP Choudhury ◽  
MA Kabir ◽  
MYA Khan ◽  
M Ershaduzzaman ◽  
...  
Keyword(s):  
Genetic Distance ◽  
Microsatellite Markers ◽  
River Basin ◽  
Genetic Relationships ◽  
Genetic Distances ◽  
Genetic Group ◽  
Sheep Population ◽  
Blood Samples ◽  
Indigenous Sheep ◽  
Pcr Product

The study was conducted to investigate the genetic relationships among indigenous sheep population of Bangladesh (Barind, Jamuna river basin, Coastal and Garole sheep) using microsatellite markers. A total of 96 blood samples were collected from adult sheep of Barind (24), Jamuna River Basin (24), Coastal (24), Garole(10) and available Chotanagpuri (10) sheep. Chotanagpuri sheep was used as an outgroup population. DNA was extracted from blood samples using QIAGEN DNA Mini extraction kit and was quantified using a nanodrop. FAO recommended 13 labeled microsatellite markers were used for polymerase chain reaction (PCR). PCR product was confirmed by 2% agarose gel electrophoresis and visualized by staining with ethidium bromide.The exact allele sizes in each primer were determined by GeneMaker V1.85 demo. Microsatellite tool kit and Dispan software package were used for calculation of allele frequency, number of alleles per locus, observed and expected heterozygosity and genetic distances (DA). The Dispan software was used to calculate inter-individual genetic distances. These distance values were used to construct an UPGMA tree. Results showed that average number of polymorphic alleles per locus varied from4 in HUJ616 to 12 in MAF70. Observed heterozygosity was also varied from 0.54±0.04 in Coastal to 0.63±0.03 in Barind sheep population. Genetic distance between Jamuna river basin and Barind was lowest (0.01) and between Garole and Costal was highest (0.17). Garoleand Chotonagpuri sheep has higher genetic distance from other three sheep populations. Phylogenetic dendogram showed that sheep of Jamuna river basin and barind were belonged to same genetic group. Whereas, coastal, garole and Nagpur sheeps were shown higher genetic distances from Jamuna river basin and coastal sheep. Considering findings of this study it may be concluded that the Barind and Jamuna river basin sheep belongs to a similar genetic group while, Garole and Coastal sheep are belonging to two distinct genetic groups. Bang. J. Anim. Sci. 2019. 48 (1): 17-22

Dizygotic monochorionic canine fetuses with blood chimaerism and suspected freemartinism

2017 ◽  
Vol 29 (2) ◽  
pp. 368 ◽  
Author(s):  
Carolynne J. Joonè ◽  
Kurt G. M. De Cramer ◽  
Johan O. Nöthling
Keyword(s):  
Genetic Analysis ◽  
Reproductive Tract ◽  
Ovarian Tissue ◽  
External Genitalia ◽  
Normal Male ◽  
Tissue Samples ◽  
Female Fetus ◽  
Blood Samples ◽  
Dna Profiles ◽  
Female External Genitalia

Two full-term canine fetuses were found to share a placenta during Caesarean section. The fetuses were of discordant gender, with apparently normal male and female external genitalia. Genetic analysis of whole-blood samples obtained from each fetus revealed identical DNA profiles, with more than two alleles detected at six loci. Subsequent genetic analysis of myocardial tissue samples revealed dissimilar DNA profiles, with at most two alleles detected per locus. Superimposition of the tissue-derived profiles matched that derived from the blood samples exactly, except for two loci failing to amplify, and hence demonstrated blood chimaerism. Dissection of the abdomen of the male fetus revealed delayed descent of the testes towards the inguinal canals. Macroscopically, the gonads, uterus and vagina were not identifiable on dissection of the female fetus, although vestigial ovarian tissue and a vagina were detected microscopically. The hypoplastic internal reproductive tract of the female fetus was suggestive of freemartinism and is believed to be the first report of this condition in the canine.

scholarly journals The Effect of Using DNA Obtained from Blood of Cattle with Genetic Chimerism on Illumina’s Beadchip Assay Performance

2014 ◽  
Vol 14 (2) ◽  
pp. 279-286 ◽  
Author(s):  
Artur Gurgul ◽  
Dominika Rubiś ◽  
Monika Bugno-Poniewierska
Keyword(s):  
Common Phenomenon ◽  
Direct Identification ◽  
Blood Samples ◽  
Precise Information ◽  
Assay Performance ◽  
Basic Parameters ◽  
Genetic Chimerism ◽  
Dna Profiles ◽  
Common User ◽  
Insight Into

Abstract Blood cell chimerism is a common phenomenon occurring in cattle coming from double or multiple parturitions and can be observed as two DNA profiles present in blood of each of twin born animals. In the era of genomics, a large number of animals is being genotyped with high throughput genotyping methods, which are giving limited insight into the performance of single markers and rather only statistical description of the results is available for a common user. This hampers the detailed analysis of the results obtained and direct identification of the causes of poorer performance of some samples. In this study we describe the influence of analysis of DNA obtained from blood samples of cattle with genetic chimerism on basic parameters of Infinium technology-based Illumina’s genotyping arrays. The results obtained may help to identify such samples, especially when no precise information about the animals’ origin is available

scholarly journals Variasi Genetik Itik Bayang Berbasis Marka Mikrosatelit Pada Lokus AY287 dan Lokus AY283

2017 ◽  
Vol 11 (2) ◽  
pp. 91 ◽  
Author(s):  
Rusfidra Rusfidra ◽  
Y. Heryandi ◽  
Jamsari Jamsari ◽  
E. Y. Rahman
Keyword(s):  
Genetic Diversity ◽  
Microsatellite Markers ◽  
Key Words ◽  
Genetic Markers ◽  
Microsatellite Loci ◽  
Gel Electrophoresis ◽  
Genetic Resource ◽  
Allele Frequencies ◽  
Blood Samples

West Sumatera Province has poultry genetic resource of local duck that potential in supply<br />eggs and duck meat. Bayang duck was set by Indonesian Agricultural Ministry in 2012 as national livestock breeds in Indonesia. Microsatellite markers are widely used as a genetic identifier because of their abundant existence, co-dominant and high polymorphic. The purpose of this study was to determine the genetic diversity of Bayang ducks based on two microsatellite loci which include AY287 and AY283. DNA substances used in the study were blood samples from 24 Bayang duck in Pesisir Selatan Regency. The isolated DNA genom from 24 blood samples of Bayang duck could be detected by gel electrophoresis. Results showed that AY287 locus has 6 alleles; allele A (108 bp), allele B (142 bp), allele C (183 bp), allele D (227 bp), allele E (272 bp) and allele F (340 bp). Both allele E and F were specific genetic markers of Bayang duck. Alleles frequencies of the AY287 locus were as follow: allele C (26,93%), allele D (19,24%), allele A (15,38%), allele B (15,38%), allele E (15,38%) and allele F (7,69%). The AY283 locus has 6 alleles consisted of allele A (230 bp), allele B (320 bp), allele C (345 bp), allele D (390 bp), allele E (450 bp) and allele F (500 bp). Allele frequencies of this marker were allele B (20,51%), allele D (20,51%), allele E (20,51%), allele A (15,39%), allele C (15,39%), and allele F (7,69%), respectively. Our finding suggest that two microsatellite markers, AY287 and AY283, were polymorphic in Bayang duck population.<br />Key words: Bayang duck, microsatellite, AY283, AY287

scholarly journals Genetic Markers for Adolescent Idiopathic Scoliosis on Chromosome 19p13.3 among Saudi Arabian Girls

2017 ◽  
Vol 11 (2) ◽  
pp. 167-173 ◽  
Author(s):  
Abdallah Ahmad Al-Othman ◽  
Mir Sadat-Ali ◽  
Ahmed Sh. Amer ◽  
Dakheel A. Al-Dakheel
Keyword(s):  
Adolescent Idiopathic Scoliosis ◽  
Idiopathic Scoliosis ◽  
Microsatellite Markers ◽  
Healthy Subjects ◽  
Genetic Locus ◽  
Saudi Arabian ◽  
Controlled Study ◽  
Blood Samples ◽  
Healthy Siblings ◽  
The University

<sec><title>Study Design</title><p>Prospective case-controlled study.</p></sec><sec><title>Purpose</title><p>This study aimed to assess genetic influence in Saudi Arabian children with adolescent idiopathic scoliosis (AIS).</p></sec><sec><title>Overview of Literature</title><p>The genetic locus linked to chromosome 19p for idiopathic scoliosis has been described. A pilot study conducted at King Fahd Hospital of the University, Al-Khobar showed that three microsatellite markers (D19S216, D19S894, and DS1034) of chromosome 19p13.3 were significant in Saudi Arabian females compared with healthy subjects.</p></sec><sec><title>Methods</title><p>A total of 100 unrelated Saudi Arabian girls treated for AIS, their parents, healthy siblings, and healthy subjects were recruited for genetic analysis of markers on chromosome 19p13.3. After informed consent was obtained from their parents, blood samples were collected and parametric and nonparametric linkage analyses were performed using GENEHUNTER ver. 2.1. Multipoint linkage analysis was used to specify an autosomal dominant trait with a gene frequency of 0.01 and an estimated penetrance of 80% at the genotypic and allelic levels.</p></sec><sec><title>Results</title><p>Five hundred blood samples were collected and analyzed for microsatellite markers (D19S216, D19S894, and DS1034) of chromosome 19p13.3. Comparison among patients, family members, and healthy subjects revealed no significant association between markers and scoliosis at the genotypic level: D19S216 (<italic>p</italic>=0.21), D19S894 (<italic>p</italic>=0.37), and DS1034 (<italic>p</italic>=0.25). However, at the allelic level, a statistically significant association was observed for marker DS1034 (<italic>p</italic>=0.008), and marker D19S216 showed significance between fathers and patients (<italic>p</italic>&lt;0.001) compared with patients and mothers. The other two markers, D19S216 (<italic>p</italic>=0.25) and D19S894 (<italic>p</italic>=0.17), showed no significant association between patients and mothers.</p></sec><sec><title>Conclusions</title><p>At the allelic level, marker DS1034 was significantly associated with AIS patients and their fathers. This allelic marker on chromosome 19p13.3 appears to be important in AIS etiology.</p></sec>

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
Keyword(s):  
Microsatellite Markers ◽  
Forensic Medicine ◽  
Tandem Repeats ◽  
Parentage Analysis ◽  
Proteinase K ◽  
Individual Identification ◽  
Statistical Parameters ◽  
Pcr Products ◽  
Hardy Weinberg Equilibrium ◽  
Forensic Parameters

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.

scholarly journals Parentage assignment and parental contribution analysis in large yellow croaker Larimichthys crocea using microsatellite markers

2012 ◽  
Vol 58 (2) ◽  
pp. 244-249 ◽  
Author(s):  
Xiande Liu ◽  
Guangtai Zhao ◽  
Zhiyong Wang ◽  
Mingyi Cai ◽  
Hua Ye ◽  
...  
Keyword(s):  
Microsatellite Markers ◽  
Small Population ◽  
Microsatellite Analysis ◽  
Large Yellow Croaker ◽  
Parentage Assignment ◽  
Larimichthys Crocea ◽  
Artificial Breeding ◽  
Resource Enhancement ◽  
Parental Contribution ◽  
Full Factorial

Abstract The large yellow croaker Larimichthys crocea is one of the most important fish species in China. To estimate the reproductive success of breeders, three independent full-factorial crosses were created and the fins of breeders and progenies were sampled for microsatellite analysis. Out of 959 offspring from three sets, 99.6% were assigned to their parents using 6–7 microsatellite markers. In all crosses, some parent pairs produced a large number of offspring and some parent pairs did not produce any offspring. The contributions of male or female parents were unequal, ranging from 1.0–89.3% across the three sets. The loss of putative Ne was 69.6% in set 1, 31.2% in set 2 and 57.6% in set 3. These results suggest that the unequal contribution of parents is universal in artificial breeding of L. crocea, especially in a small population, and this should be taken into account in hatcheries or when releasing animals for resource enhancement [Current Zoology 58 (2): 244–249, 2012].

Twelve microsatellite markers for characterization of Plasmodium falciparum from finger-prick blood samples

Parasitology ◽  
1999 ◽  
Vol 119 (2) ◽  
pp. 113-125 ◽  
Author(s):  
T. J. C. ANDERSON ◽  
XIN-ZHUAN SU ◽  
M. BOCKARIE ◽  
M. LAGOG ◽  
K. P. DAY
Keyword(s):  
Plasmodium Falciparum ◽  
Microsatellite Markers ◽  
Blood Samples ◽  
Field Samples ◽  
Finger Prick ◽  
Pcr Products ◽  
Malaria Species ◽  
Carry Over ◽  
The Village

Multiple, selectively neutral genetic markers are the most appropriate tools for analysis of parasite population structure and epidemiology, but yet existing methods for characterization of malaria field samples utilize a limited number of antigen encoding genes, which appear to be under strong selection. We describe protocols for characterization of 12 microsatellite markers from finger-prick blood samples infected with Plasmodium falciparum. A two-step, heminested strategy was used to amplify all loci, and products were visualized by fluorescent end-labelling of internal primers. This procedure allows amplification from low levels of template, while eliminating the problem of spurious products due to primer carry over from the primary round of PCR. The loci can be conveniently multiplexed, while accurate sizing and quantification of PCR products can be automated using the GENOTYPER software. The primers do not amplify co-infecting malaria species such as P. vivax and P. malariae. To demonstrate the utility of these markers, we characterized 57 infected finger-prick blood samples from the village of Mebat in Papua New Guinea for all 12 loci, and all samples were genotyped a second time to measure reproducibility. Numbers of alleles per locus range from 4 to 10 in this population, while heterozygosities range from 0·21 to 0·87. Reproducibility (measured as concordance between predominant alleles detected in replicate samples) ranged from 92 to 98% for the 12 loci. The composition of PCR products from infections containing multiple malaria clones could also be defined using strict criteria and scored in a highly repeatable manner.

scholarly journals Implications for the use of horse hair roots as a DNA source for microsatellite typing

2011 ◽  
Vol 50 (No. 11) ◽  
pp. 499-502 ◽  
Author(s):  
T. Ząbek ◽  
A. Radko ◽  
E. Słota
Keyword(s):  
Dna Typing ◽  
Hair Follicles ◽  
Proteinase K ◽  
Digestion Method ◽  
Preparation Methods ◽  
Dna Preparation ◽  
Hair Roots ◽  
Horse Hair ◽  
Dna Profiles ◽  
Parentage Control

Hair roots are a very attractive source of DNA for microsatellite-based parentage control of breeding animals. However, unlike blood samples, irregular DNA typing results have been observed in assays utilizing hair follicles. The amount of starting material and DNA preparation method are the crucial factors. In order to improve DNA typing results for horse hair roots, two quick preparation methods and additional purification steps were evaluated. PCR efficiency for each approach was expressed as percentage of samples with complete DNA profiles for 12 horse microsatellites. The lowest percentage (22%) of complete DNA profiles was obtained for samples prepared by the proteinase K digestion method. The best genotyping results (94%) were achieved after phenol-chloroform extraction of DNA from samples prepared by the proteinase K digestion method. Direct cleanup of DNA samples with an ethanol-sodium acetate mixture gave comparably good results of microsatellite genotyping (91%). DNA preparation from hair roots with proteinase K digestion followed by DNA purification with ethanol was chosen as the most efficient approach for horse DNA typing under parentage testing. &nbsp; &nbsp;

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