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PMID: 10933851 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Anchor polymerase chain reaction display: a high-throughput method to resolve, score, and isolate dimorphic genetic markers based on interspersed repetitive DNA elements.

Analytical biochemistry ·Vol. 284 ·No. 1 ·2000-08-15 ·Pages 19-28

Ayyadevara S, Thaden JJ, Shmookler Reis RJ

Abstract

Genes which confer a disease when mutated, or for which population variability contributes to a quantitative trait such as longevity or disease susceptibility, can be localized in the genetic map by use of an appropriately dense set of polymorphic DNA markers. Here we describe an anchor PCR method for high-throughput genotyping, which can be used to amplify the DNA segments flanking an interspersed repetitive sequence such as a transposon, and to limit the number of product bands per reaction to facilitate marker resolution. We used this method to amplify and display DNA fragments flanking the Tc1 transposable elements from different strains of the nematode Caenorhabditis elegans, varying widely in insert number, and to analyze marker segregation in recombinant inbred lines generated from an interstrain cross. Since essentially all eukaryotic genomes contain abundant interspersed repeat families, many of which are dimorphic (for presence or absence of specific elements) among populations, this method can be used for rapid genotyping and fine-scale chromosomal mapping in many species, including those for which extensive mapping and sequencing data do not yet exist.

MeSH Terms
Animals Caenorhabditis elegans/genetics Chromosome Mapping/methods DNA/metabolism DNA Transposable Elements Genetic Markers Genotype Polymerase Chain Reaction/methods Quantitative Trait, Heritable Repetitive Sequences, Nucleic Acid
Chemicals
DNA Transposable Elements Genetic Markers DNA
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Ayyadevara S
Department of Biochemistry and Molecular Biology, University of Arkansas for Medical Sciences, Little Rock, Arkansas 72205, USA.
Thaden J J
Shmookler Reis R J
Article Info
Journal
Analytical biochemistry
Abbr.
Anal Biochem
ISSN
0003-2697
Published
2000-08-15
Pages
19-28
Language
English
Region
United States
NLM ID
0370535
Subset
IM
Grants
NIA NIH HHS · P01-AG13918 · United States
NIA NIH HHS · R01-AG09413 · United States
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