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Journal of Heredity Advance Access published online on December 12, 2008
Journal of Heredity, doi:10.1093/jhered/esn108
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Published by Oxford University Press 2008.

Brief Communication

AFLP Fragment Isolation Technique as a Method to Produce Random Sequences for Single Nucleotide Polymorphism Discovery in the Green Turtle, Chelonia mydas

Suzanne E. Roden, Peter H. Dutton, and Phillip A. Morin

From the National Marine Fisheries Service, Southwest Fisheries Science Center, 8604 La Jolla Shores Drive, La Jolla, CA 92037 (Roden, Dutton, and Morin); the University of San Diego, Marine Science Department, 5998 Alcalà Park, San Diego, CA 92110 (Roden)

Address correspondence to Suzanne E. Roden at the address above, or e-mail: suzanne.roden{at}noaa.gov.

The green sea turtle, Chelonia mydas, was used as a case study for single nucleotide polymorphism (SNP) discovery in a species that has little genetic sequence information available. As green turtles have a complex population structure, additional nuclear markers other than microsatellites could add to our understanding of their complex life history. Amplified fragment length polymorphism technique was used to generate sets of random fragments of genomic DNA, which were then electrophoretically separated with precast gels, stained with SYBR green, excised, and directly sequenced. It was possible to perform this method without the use of polyacrylamide gels, radioactive or fluorescent labeled primers, or hybridization methods, reducing the time, expense, and safety hazards of SNP discovery. Within 13 loci, 2547 base pairs were screened, resulting in the discovery of 35 SNPs. Using this method, it was possible to yield a sufficient number of loci to screen for SNP markers without the availability of prior sequence information.

Key Words: AFLP • Chelonia mydas • green turtle • single nucleotide polymorphism • SNP

Corresponding Editor: Brian Bowen

Received August 26, 2008
Revised November 14, 2008
Accepted November 19, 2008


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