Differentiation of Anopheles minimus Species Complex by RAPD-PCR Technique

Abstract

Amplification of random regions of genomic DNA using 10-base primers in the randomamplified polymorphic DNA polymerase chain reaction (RAPD-PCR) was used to differentiate Anopheles minimus A and An. minimus C. Genomic DNA was extracted from individual mosquitoes of An. minimus A and An. minimus C and amplified in PCR reactions using single primers of arbitrary nucleotide sequence. Fifteen different commercially available primers (Operon oligonucleotides kit M from Operon Technologies, Inc.), six primers were selected on the basis of presence or absence of the bands of A and C. They gave 8 different amplified DNA fragments of these two species of An. minimus. The primers revealed only species A are OPM8 and OPM13 at the 0.8 and 2.15 and 2, while both species A and C were in OPM17 at 0.8 Kb revealed A, at the 0.55 and 1.5 Kb revealed C. OPM 12 gave a 0.5 Kb DNA fragment in A while 0.4 Kb in C. OPM15 showed C at 0.7 Kb. These findings indicated that An. minimus species A and C can be differentiated by RAPD-PCR technique.