2.2. Microsatellite genotyping of samples
Thirty A. stephensi mosquitoes from each population were genotyped
at 13 microsatellite loci developed by Veradi et al. (2002)
(Table S1). Each pair of microsatellite markers were labeled by 6-FAM.
Multiplex PCR was carried out by grouping the microsatellite markers
according to their allele size range and annealing temperature. The
combinations used were D8T & E7; H2II & E12; B2N & F10; H1, A7 &
A10; G1, B2 &MSH1; B1. The reaction conditions were, initial heat activation
at 95 °C for 15 min, denaturation at 94 °C for 30 s, annealing at
57–60 °C depending on combination used and extension at 72 °C for
90 s. The numbers of cycles used were 30–45. Genotyping was done
by ABI 3730xl genetic analyzer and the data was analyzed by the
Genemapper 4.0 software.