A set of canine interrepeat sequence PCR markers for high-throughput genotyping.
One hundred and sixteen interspersed repetitive DNA sequence (IRS)-PCR markers have been developed and characterized from Canis familiaris for high-throughput filter-based genotyping. We present a detailed analysis of markers produced by amplification using primers directed to the conserved regions of the C. familiaris short interspersed nuclear element (Can-SINE). The majority of IRS-PCR markers developed were moderately to highly polymorphic with mean heterozygosity (HET) and polymorphism information content (PIC) values of approximately 0.6. The HET value for 22.3% of the markers exceeded 0.7. We also demonstrate that sequence variation of Can-SINEs between breeds is significant and also represents a rich source of polymorphisms. Mapping of 73 of the markers to the existing integrated linkage-radiation hybrid map enriches the map as well as establishes the utility of the markers. The significance and utility of this new class of IRS-PCR Can-SINE-based markers for high-throughput genotyping is discussed. This method can also be extended to other species that are currently map-poor but have a sufficiently high density of SINEs to allow IRS-PCR.
Das
M
M
Department of Biochemistry, Department of Oncology, McGill University, Montreal, Quebec, Canada H3G 1Y6.
Sakul
H
H
Kong
J
J
Acland
G M
GM
Pelletier
J
J
D000818:Animals; D017124:Conserved Sequence; D004285:Dogs / Q000235:genetics*; D008040:Genetic Linkage; D005819:Genetic Markers / Q000235:genetics; D005838:Genotype; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011110:Polymorphism, Genetic / Q000235:genetics; D018807:Polymorphism, Single-Stranded Conformational; D021101:Radiation Hybrid Mapping / Q000662:veterinary; D020086:Short Interspersed Nucleotide Elements / Q000235:genetics*
D005819:Genetic Markers