SIMPLE SEQUENCE REPEAT-BASED SINGLE PRIMER AMPLIFICATION OF GENOMIC DNA IN RANDOM BRED POPULATIONS OF TURKEYS AND CHICKENS

The ubiquity and polymorphism of simple sequence repeats (SSR) or micr osatellites have made them the DNA markers of choice for linkage, dive rsity and phylogenetic studies in eukaryotes. In this study, we invest igated the utility of a SSR-based single primer amplification procedur e (SPARs) for developing DNA markers in turkeys and chickens. The fift een primers used to evaluate the average number, size, and polymorphis m of DNA fragments within SPARs patterns consisted of one di-, six tri - and eight tetra-nucleotide repeats. Genomic DNA samples from a total of 60 and 40 random bred turkeys and chickens, respectively, were use d. In addition, turkey reference families developed from a wild x Comm ercial Orlopp-line reciprocal cross was included in the analyses. Cycl ing conditions for the polymerase chain reaction were optimized for ea ch primer by testing for amplification using annealing temperatures of 35, 45 and 55 degrees C. Molecular weight, number and level of polymo rphism of the amplified DNA fragments were primer dependent. A total o f 66 fragments in turkeys and 83 in chickens was amplified, of which o nly 11 and 15% in turkeys and chickens, respectively, were polymorphic . Amplified fragments in both species ranged in molecular weight from 150 to 2500 base pairs. About 90% of the amplified fragments were spec ies-specific in turkeys and in chickens. The low level of polymorphic fragments in the populations analyzed suggests that SPARs may be an in efficient method for the development of markers in both turkeys and ch ickens.