Cytochrome Oxidase I (COI) sequences were PCR-amplified using the standard barcoding primers LCO1490 (3'-GGTCAACAAATCATAAAGAT ATTGG-5') and HCO2198 (5'- TAAACTTCAGGGTGACCAAAAAATCA-3'). PCR was performed with a Eppendorf thermocycler in 30 ul reactions containing 1.5 ul of each primer (10pmol), 20.5 ul water, 3 ul ExTAQ buffer, 2.4 ul  Ex Taq dNTP mix, 1 unit of Taq polymerase (Takara Ex Taq), and 1 ul DNA. The reaction took place under the following cycling conditions: 2 min of denaturation at 94ºC, followed by 35 cycles of denaturation at 94ºC for 20 sec., annealing at 50ºC for 20 sec., and primer extension at 72ºC for 1.35 min, and a final extension at 72ºC for 5 min. The PCR products were purified and sequenced on an Applied Biosystems 3730XL DNA Analyzer at the University of Arizona.

Users wishing to identify the pink bollworm molecularly should follow the protocol above and submit the resulting sequence to a BLAST search to compare the resulting sequence to that of the pink bollworm.