Rapid isolation of CA microsatellites from the tilapia genome

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 a...

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Bibliographic Details
Published in:Animal genetics 2002-04, Vol.33 (2), p.140-144
Main Authors: Carleton, K. L., Streelman, J. T., Lee, B.-Y., Garnhart, N., Kidd, M., Kocher, T. D.
Format: Article
Language:English
Subjects:
Animals
CA repeat
Cichlids - genetics
Dinucleotide Repeats
DNA Primers
Gene Library
Genetic Markers
Genome
linkage mapping
marker generation
microsatellite
Oreochromis aureus
Oreochromis niloticus
Polymerase Chain Reaction
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Summary: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 × 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.
ISSN:0268-9146
1365-2052
DOI:10.1046/j.1365-2052.2002.00817.x