Single primer amplification of avian genomic DNA detects polymorphic LOCI

Abstract

DNA amplification reactions using long, non-random, single oligonucleotide primers with thermostable DNA polymerase and avian genomic DNA generate unique, reproducible multiband patterns of DNA fragments. These DNA fragments are produced from amplification reactions using single primers of at least twentyfive bases in length which were derived from the chicken α A -globin gene. The patterns of amplified DNA fragments were polymorphic among individual chickens. Within families, polymorphic band patterns were found to be heritable, as the DNA fragments of progeny were present in at least one of the parents. In a conventional polymerase chain reaction containing a pair of primers, the relative concentrations of the primers can be manipulated to produce either a discrete two primer product or a multiband pattern identical to the single primer products. In addition, the number of bands in the multiband pattern increases as the stringency of the annealing step of the DNA amplification is decreased. The products are believed to result from recognition of a family of related sequences in the genome. This single primer amplification method has general application in the establishment of molecular relatedness, discovery of new genetic markers, resolution of questions of paternity, identification of individual organisms, and measurement of biodiversity within and among populations.

MAAP: a versatile and universal tool for genome analysis

“…Amplification of genomes of high complexity (soybean and human) with primers of more than 8 nt in length and of low complexity (bacteria and fungi) with primers of more than 7 nt in length produce many more products than expected (Table 2; Fig. Moreover, primers of length comparable to those used in the PCR (20-60 nt in length) using AP-PCR [86] or RAPD [1,21,28,61,63] techniques produced fingerprints from bacterial, fungal, animal and plant species. For example, decamer primers produced up to 60 amplification products when DNA from a caucasian human was amplified [13].…”

Genetic Markers, Map Construction, and Their Application in Plant Breeding

“…RAPDs. Of three similar, single-primer, PCR-based technologies, random amplified polymorphic DNA (RAPD; Williams et al, 1990), DNA amplification fingerprinting (DAF;Caetano-Anolles et al, 1991), and arbitrary-primed PCR, (AP-PCR; Owen and Uyeda, 1991;Welsh and McClelland, 1990), RAPDs have been used most widely for map construction and linkage analysis (Reiter et al, 1992) (Fig. 1).…”