ABSTRACT
Identification of genetic markers that can be used to discriminate among geographic populations of Ceratitis capitata is a high priority for analysis of global spread of this pest. An earlier report demonstrated the utility of mitochondrial DNA (mtDNA) restriction enzyme techniques in separating populations into two major categories on the basis of variation in the number of fragments generated by the enzymes EcoRV and XbaI. Relative positions of some mitochondrial genes were determined by a combination of polymerase chain reaction (PCR), sequence analysis, and restriction enzyme analysis. PCR amplification of a 2.07 kbp containing the EcoRV variable site was performed using primers in the ND4 and cytochrome b genes and mtDNA extracted from Guatemalan flies. This amplified fragment was then cloned into the pCRII vector using the TA cloning kit. Single individuals were analyzed using both our original technique and the new PCR technique to confirm the accuracy of haplotype designation.
