title: Rapid isolation of CA microsatellites from the tilapia genome.
abstract: We have developed (CA)n microsatellite markers for the cichlid fish, Oreochromis niloticus using a variation of the hybrid capture method. The resulting genomic library was highly enriched in repetitive DNA with 96% of clones containing CA repeats. The number of repeats ranged from four to 45 with an average of 19. Two-thirds of the sequenced clones had 12 or more repeats and sufficient flanking sequence to design primers. The resulting markers were tested in an F2 cross of O. niloticus x O. aureus. Nearly 90% of the markers amplified in this cross and 74% of these were informative. This work demonstrates the importance of minimizing the number of polymerase chain reaction (PCR) amplification cycles before and after the enrichment steps to reduce PCR recombination and the generation of chimaeric clones.
authors.affiliation: Hubbard Center for Genome Studies, University of New Hampshire, Durham, NH 03824, UK. karen.carleton@unh.edu
authors.forename: K L
authors.initials: KL
authors.lastname: Carleton
authors.forename: J T
authors.initials: JT
authors.lastname: Streelman
authors.forename: B-Y
authors.initials: BY
authors.lastname: Lee
authors.forename: N
authors.initials: N
authors.lastname: Garnhart
authors.forename: M
authors.initials: M
authors.lastname: Kidd
authors.forename: T D
authors.initials: TD
authors.lastname: Kocher
mesh_terms: D000818:Animals; D023681:Cichlids / Q000235:genetics*; D017931:DNA Primers; D018900:Dinucleotide Repeats; D015723:Gene Library; D005819:Genetic Markers; D016678:Genome; D016133:Polymerase Chain Reaction
chemical_list: D017931:DNA Primers; D005819:Genetic Markers