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Rapid isolation of CA microsatellites from the tilapia genome
Author(s) -
Carleton K. L.,
Streelman J. T.,
Lee B.Y.,
Garnhart N.,
Kidd M.,
Kocher T. D.
Publication year - 2002
Publication title -
animal genetics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.756
H-Index - 81
eISSN - 1365-2052
pISSN - 0268-9146
DOI - 10.1046/j.1365-2052.2002.00817.x
Subject(s) - biology , microsatellite , polymerase chain reaction , genome , genetics , tilapia , oreochromis , cichlid , genomic dna , genomic library , microbiology and biotechnology , dna , gene , fish <actinopterygii> , base sequence , allele , fishery
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 F 2 cross of O. niloticus × 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.