5' and 3' SINE-PCR allows genotyping of pig families without cloning and sequencing steps.
A microsatellite-containing clone, isolated from a pig Chromosome (Chr) 1-specific library was characterized by sequencing and computer analysis. The (CA)17 microsatellite motif was located at the 3' end of a short interspersed element (SINE) sequence at the position normally occupied by the oligo (A) stretch. Further computer analysis indicated that 12% of published pig SINE sequences contain dinucleotide repeat motifs adjacent to their 3' ends. By performing PCR with a single SINE primer in combination with a panel of arbitrarily selected unique primers, we have demonstrated that, as in human, polymorphisms can be detected and typed in pig family DNAs. A large number of SINE primer x unique primer combinations have been screened for the ability to detect polymorphisms in pig reference family DNAs. This approach does not require prior sequence information other than that of the pig SINE. We have also found polymorphisms at the 5' ends of pig SINE sequences by similar methods, but with a primer facing out to the 5' end of the SINE.
Miller
J R
JR
Agricultural and Food Research Council Institute of Animal Physiology and Genetics Research Cambridge Research Station, UK.
Archibald
A L
AL
D000818:Animals; D001483:Base Sequence; D004276:DNA, Satellite / Q000235:genetics; D005069:Evaluation Studies as Topic; D005260:Female; D015723:Gene Library; D005819:Genetic Markers; D005838:Genotype; D008297:Male; D008969:Molecular Sequence Data; D010375:Pedigree; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D013552:Swine / Q000235:genetics*
D004276:DNA, Satellite; D005819:Genetic Markers