Cloning of highly polymorphic microsatellites in the horse.
Abstract: We have isolated equine microsatellites by screening a genomic library with (TG)n and (TC)n probes. TG microsatellites were found to be more abundant than TC repeats, with an estimated frequency of one per 100,000bp. Sequence analysis of eight TG-positive clones revealed varying structures of the repeat regions; perfect stretches of TG repeats, imperfect stretches of TG repeats and compound regions of TG and TC repeats. Five loci were analysed by PCR and showed extensive polymorphism; three to seven alleles and heterozygosities of 0.40-0.76 were observed when screening 20-30 unrelated individuals. The high degree of polymorphism, their abundance and the possibility of automating the typing procedure make these loci ideal for standardized paternity testing in the horse. Furthermore, we demonstrate that single hairs can be used as starting material for the PCR analysis.
Publication Date: 1992-01-01 PubMed ID: 1443772DOI: 10.1111/j.1365-2052.1992.tb00032.xGoogle Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article discusses the successful cloning of highly variable microsatellites in horses, which have significant potential for usage in standardized paternity testing due to their high degree of polymorphism. The study also demonstrated the possibility of using single hairs as base material in the process of Polymerase Chain Reaction (PCR) analysis.
Isolation of Equine Microsatellites
- The researchers isolated equine microsatellites by screening a genomic library with (TG)n and (TC)n probes.
- Microsatellites with TG repeat structure were found more abundantly than those with TC repeats, with a frequency of one in every 100,000 base pairs.
Sequence Analysis of Clones
- After conducting a sequence analysis on eight TG-positive clones, the researchers observed that the repeat regions had various structures.
- These structures ranged from perfect stretches of TG repeats to imperfect stretches and compound regions of both TG and TC repeats.
PCR Analysis and Polymorphism
- PCR analysis was performed on five selected loci, revealing extensive polymorphism.
- When screening 20-30 unrelated individuals, the degree of heterozygosity ranged from 0.40 to 0.76, with each locus having between three to seven alleles.
- This high level of polymorphism, along with their abundance and the potential for automation, makes these loci highly suitable for standardized paternity testing in horses.
Use of Single Hairs in PCR Analysis
- The researchers were successfully able to utilize single hairs as initial material for the PCR analysis.
- This indicates that the method can be practical, as it proves that it is possible to conduct this analysis with minimalistic starting material.
Cite This Article
APA
Ellegren H, Johansson M, Sandberg K, Andersson L.
(1992).
Cloning of highly polymorphic microsatellites in the horse.
Anim Genet, 23(2), 133-142.
https://doi.org/10.1111/j.1365-2052.1992.tb00032.x Publication
Researcher Affiliations
- Department of Animal Breeding and Genetics, Swedish University of Agricultural Sciences, Uppsala.
MeSH Terms
- Animals
- Base Sequence
- Cloning, Molecular
- DNA
- DNA, Satellite / genetics
- Hair
- Horses / genetics
- Molecular Sequence Data
- Polymerase Chain Reaction / veterinary
- Polymorphism, Genetic
Citations
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