The thief ant, Solenopsis molesta (Say), a common nuisance species found throughout the United States is genetically related to red imported fire ants, S. invicta Buren. Therefore, its identification at the molecular level is very important. The deoxyribonucleic acid (DNA) barcoding, a recent technique was used to identify thief ant complex at species and subspecies levels using a short DNA sequence from the cytochrome oxidase subunit 1 (COI) mitochondrial region. The DNA from thief ants collected from 9 states was extracted using Qiagen's Gentra PUREGENE® DNA Isolation Kit. The polymerase chain reactions (PCR) were run on the extracted DNA to amplify partial sequence of COI using primers Lep-F1 (forward) and Lep-R1 (reverse). The resulting DNA products were concentrated, purified and sequenced. The 600 bp sequences of the COI generated were submitted to GenBank that issued accessions numbers from HM179641 to HM179653. The sequences associated with these accession numbers were used as DNA barcodes for distinguishing species and subspecies. Based on this molecular analysis, thief ants collected from New York, Indiana and 1 location in Nebraska were separated in 1 group as S. molesta validiuscula (Emery) and another with ants from Louisiana identified as S. carolinensis (Forel). The third group was comprised of ants from South Dakota, Washington, New Jersey, Tennessee, Kansas and 2 other locations in Nebraska was identified as S. molesta molesta (Say).
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