• Title/Summary/Keyword: Microsatellite

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Development of a Multiplex PCR System for Microsatellite Genotyping of the Sea Cucumber Stichopus japonicus (해삼(Stichopus japonicus)의 microsatellite 유전자형 분석을 위한 multiplex PCR 시스템 개발)

  • Sim, Yong-Teak;Lee, Chul-Sang
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.6
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    • pp.806-811
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    • 2017
  • A multiplex PCR system comprising 14 microsatellite markers was developed for genotyping analysis of the sea cucumber Stichopus japonicus. A total of 286 samples were used to evaluate genetic polymorphisms and forensic parameters of the microsatellite loci. In a single PCR reaction, all 14 loci were uniformly amplified and a total of 269 alleles were identified. The AJ19024 locus had the largest number of alleles (46), and its discriminatory power and exclusion power were 0.99 and 0.76, respectively. The fewest alleles (8) were present at the Psj2575 locus, which provided the lowest discriminatory power (0.81) and exclusion power (0.20). The mean number of alleles, mean heterozygosity, mean discrimination power and mean exclusion power per locus were 19.21, 0.70, 0.93, and 0.46, respectively. The combined matching probability for the 14 loci was $9.64{\times}10^{-19}$, and the combined power of exclusion was 0.999995. Thus, the forensic parameters evaluated in the present study demonstrated the utility of our multiplex PCR system for biological tracing methods, such as individual identification and paternity testing, in the sea cucumber.

Evaluation of Genetic Relationship and Fingerprinting of Rice Varieties using Microsatellite and RAPD Markers

  • Soo- Jin, Kwon;Sang-Nag, Ahn;Hae-Chune, Choi;Huhn-Pal, Moon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.44 no.2
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    • pp.112-116
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    • 1999
  • Genetic diversity of 31 rice varieties including 25 japonica and 6 indica varieties was evaluated using a combination of 19 microsatellite or simple sequence repeats (SSRs) and 28 random decamer oligonucle-otide primers. All 19 microsatellite primer sets representing 19 loci in the rice genome showed polymorphisms among the 31 varieties and revealed 91 alleles with an average of 4.80 bands per primer. Also all 28 random decamer primers used were informative and generated 114 non-redundant bands with a mean of 4.07 bands. Microsatellite markers detected higher number of alleles than random primers .although the mean difference was not statistically significant. A cluster analysis based on Nei's genetic distances calculated from the 205 bands resolved the 31 varieties into two major groups that correspond to indica and japonica subspecies, which is consistent with the genealogical information. As few as six random decamer primers or a combination of one microsatellite and four random decamer primers were sufficient to uniquely differentiate all 31 varieties. These combinations would be potentially useful in rice variety protection and identification considering that 25 out of 31 varieties used in this study are japonica rices with high grain quality and have close make up.

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Correlations between Heterozygosity at Microsatellite Loci, Mean d2 and Body Weight in a Chinese Native Chicken

  • Liu, G.Q.;Jiang, X.P.;Wang, J.Y.;Wang, Z.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.12
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    • pp.1671-1677
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    • 2006
  • A total of two hundred twenty eight half-sib chickens were scored for allele size at 20 microsatellite loci to estimate individual heterozygosity and mean $d^2$. The averages of microsatellite heterozygosity, allele per locus and mean $d^2$ were 0.39, 3.6 and 49, respectively. The body weight was measured biweekly from birth to twelve weeks of age. Gompertz function was assumed to simulate body weight and to estimate the growth model parameters. Due to sex effect on body weight, the regression of body weight on heterozygosity as well as on mean $d^2$ in males and females was analyzed separately in the present study. Positive correlations were found between microsatellite heterozygosity and body weight in males and females (p<0.05). Positive correlation also observed between individual heterozygosity and simulated maximum daily gain estimated from Gompertz function in female chickens (p<0.05). There were no significant correlations between mean $d^2$ and body weight. The results suggest that local effect hypothesis could explain the correlations between heterozygosity and fitness-related traits in the domesticated chicken population, rather than the general effect hypothesis does.

Evaluation of Genetic Variability in Kenkatha Cattle by Microsatellite Markers

  • Pandey, A.K.;Sharma, Rekha;Singh, Yatender;Prakash, B.;Ahlawat, S.P.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.12
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    • pp.1685-1690
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    • 2006
  • Kenkatha cattle, a draft purpose breed, which can survive in a harsh environment on low quality forage, was explored genetically exploiting FAO-suggested microsatellite markers. The microsatellite genotypes were derived by means of the polymerase chain reaction (PCR) followed by electrophoretic separation in agarose gels. The PCR amplicons were visualized by silver staining. The allelic as well as genotypic frequencies, heterozygosities and gene diversity were estimated using standard techniques. A total of 125 alleles was distinguished by the 21 microsatellite markers investigated. All the microsatellites were highly polymorphic with mean allelic number of 5.95${\pm}$1.9 (ranging from 3-10 per locus). The observed heterozygosity in the population ranged between 0.250 and 0.826 with a mean of 0.540${\pm}$0.171, signifying considerable genetic variation. Bottleneck was examined assuming all three mutation models which showed that the population has not experienced bottleneck in recent past. The population displayed a heterozygote deficit of 21.4%. The study suggests that the breed needs to be conserved by providing purebred animals in the breeding tract.

Development of Polymorphic Microsatellite Markers Suitable for Genetic Linkage Mapping of Olive Flounder Paralichthys olivaceus

  • Kim, Woo-Jin;Shin, Eun-Ha;Kong, Hee Jeong;Nam, Bo-Hye;Kim, Young-Ok;Jung, Hyungtaek;An, Cheul Min
    • Fisheries and Aquatic Sciences
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    • v.16 no.4
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    • pp.303-309
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    • 2013
  • Microsatellite markers are important for gene mapping and for marker-assisted selection. Sixty-five polymorphic microsatellite markers were developed with an enriched partial genomic library from olive flounder Paralichthys olivaceus an important commercial fish species in Korea. The variability of these markers was tested in 30 individuals collected from the East Sea (Korea). The number of alleles for each locus ranged from 2 to 33 (mean, 17.1). Observed and expected heterozygosity as well as polymorphism information content varied from 0.313 to 1.000 (mean, 0.788), from 0.323 to 0.977 (mean, 0.820), and from 0.277 to 0.960 (mean, 0.787), respectively. Nine loci showed significant deviation from the Hardy-Weinberg equilibrium after sequential Bonferroni correction. Analysis with MICROCHECKER suggested the presence of null alleles at five of these loci with estimated null allele frequencies of 0.126-0.285. These new microsatellite markers from genomic libraries will be useful for constructing a P. olivaceus linkage map.

Beagle dogs parentage testing by using 22 ISAG microsatellite markers

  • Ji, Hye-jung;Kim, Eun-hee;Lee, Kyoung-kap;Kang, Tae-young;Lee, Joo-myoung;Shin, Hyoung-doo;Kim, Lyoung-hyo;Yun, Young-min
    • Korean Journal of Veterinary Research
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    • v.47 no.4
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    • pp.457-460
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    • 2007
  • The objective of the study was to establish routine parentage testing system in Beagle dogs using 22 ISAG (International Society for Animal Genetics) canine microsatellite markers (2005). Blood collections were obtained from a mother dog, 4 candidate father dogs and 3 offspring (n = 8). Genomic DNA samples were extracted from 8 Beagle dogs blood for PCR analysis. PCR products for the allele were analyzed by ABI 3130 DNA Sequencer and GeneScan (Ver 3.0) analysis and Genotyper (Ver. 2.1) software. The genetic relationship of mother and 3 offspring as well as one father dog among 4 candidate father dogs was confirmed by microsatellite allele analysis. The results of locus for amelogenin, which was designed for sexing, were matching with real gender among 8 Beagle dogs (female; 217/217 homozygosity, male; 179/217 heterozygosity). Twenty two ISAG microsatellite markers are useful the parentage test of Beagle dogs. In addition, amelogenin is an applicable marker to detecting real sex in dogs.

Development and characterization of 21 microsatellite markers in Daphne kiusiana, an evergreen broad-leaved shrub endemic to Korea and Japan

  • Lee, Jung-Hyun;Cho, Won-Bum;Yang, Sungyu;Han, Eun-Kyeong;Lyu, Eun-Seo;Kim, Wook Jin;Moon, Byeong Cheol;Choi, Goya
    • Korean Journal of Plant Taxonomy
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    • v.47 no.1
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    • pp.6-10
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    • 2017
  • Microsatellite markers were isolated for Daphne kiusiana var. kiusiana (Thymelaeaceae), an evergreen broad-leaved shrub endemic to Korea and Japan. Because its populations in Jeju Island are morphologically controversial, and consistently threatened by anthropogenic pressures, taxonomic delimitation and conservation effort are required at the genetic level. We developed 21 polymorphic microsatellite loci from Next Generation Sequencing data. The primer set included di-, tri-, and tetra-nucleotide repeats. Variability in the markers was tested for 80 individuals of D. kiusiana from three natural populations in Jeju Island and Japan. Among the 21 loci, three were unavailable for population JKJU of Japan. The Neighbor-Joining tree based on microsatellite markers described here classified the three populations into two groups according to geographical or morphological traits. These will be a powerful genetics tool for determining the taxonomic boundary and establishing suitable conservation strategies for D. kiusiana in Jeju Island.

Genetic structure of Larimichthys polyactis (Pisces: Sciaenidae) in the Yellow and East China Seas inferred from microsatellite and mitochondrial DNA analyses

  • Kim, Jin-Koo;Min, Gi-Sik;Yoon, Moon-Geun;Kim, Yeong-Hye;Choi, Jung-Hwa;Oh, Taeg-Yun;Ni, Yong
    • Animal cells and systems
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    • v.16 no.4
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    • pp.313-320
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    • 2012
  • Genetic variation was surveyed at four microsatellite loci and 1416 base pairs (bp) of the mitochondrial DNA (mtDNA) cytochrome c oxidase I gene (COI) to clarify the genetic structure of the small yellow croaker, Larimichthys polyactis, in the Yellow and East China Seas, especially regarding four provisional populations, (one Korean and three Chinese populations). Based on microsatellite DNA variations, the estimated expected heterozygosity ($H_E$) in each population ranged from 0.776 to 0.947. The microsatellite pairwise $F_{ST}$ estimates showed no significant genetic differentiation between the populations. MtDNA variations also indicated no genetic structure in L. polyactis, but very high variability. The absence of genetic differentiation among and within populations of L. polyactis may either result from the random migration of the adult or the passive dispersal of the eggs and larvae.

Microsatellite analysis of 20 mulberry varieties preserved in Korea

  • Chan Young, Jeong;Sang Kuk, Kang;Nam-Suk, Kim;Ik Seob, Cha;Seong-Wan, Kim;Jong Woo, Park;Kee-Young, Kim
    • International Journal of Industrial Entomology and Biomaterials
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    • v.45 no.2
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    • pp.49-55
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    • 2022
  • A total of 20 mulberry varieties preserved in Korea were typed for eight polymorphic microsatellite loci. We obtained 6 to 15 alleles per locus with an average value of 10.6, per-locus observed heterozygosity ranging from 0.35 to 1.00, and per-locus polymorphic information content (PIC) ranging from 0.61 to 0.87, indicating that most loci are highly variable. Phylogenetic analysis using the eight microsatellite loci was sufficiently suitable for classifying 20 mulberry varieties preserved in Korea. A total of 160 variety-specific apomorphic alleles were obtained from eight loci discriminated 20 mulberry varieties. These variety-specific alleles from this analysis are expected to be useful for the discrimination of other mulberry varieties. Furthermore, a substantial number of homozygote loci, represented by 60 among 180 alleles in eight loci were found. These results collectively suggest that these microsatellite locus primers are potentially crucial molecular markers for the eventual classification of mulberry varieties that are preserved as hundreds in Korea.

DNA Marker Mining of BMS1167 Microsatellite Locus in Hanwoo Chromosome 17

  • Lee, Jea-Young;Lee, Yong-Won;Kwon, Jae-Chul
    • Journal of the Korean Data and Information Science Society
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    • v.17 no.2
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    • pp.325-333
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    • 2006
  • We describe tests for detecting and locating quantitative traits loci (QTL) for traits in Hanwoo. Lod scores and a permutation test have been described. From results of a permutation test to detect QTL, we select major DNA markers of BMS1167 microsatellite locus in Hanwoo chromosome 17 for further analysis. K-means clustering analysis applied to four traits and eight DNA markers in BMS1167 resulted in three cluster groups. We conclude that the major DNA markers of BMS1167 microsatellite locus in Hanwoo chromosome 17 are markers 100bp, 108bp and 110bp.

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