Anopheles stephensi

Anopheles stephensi View in CoL mosquito collection and P47Rec sequence eXtraction

Anopheles stephensi DNA View in CoL generated from previous studies [ 6, 43] were used for the analyses described below. Tese source specimens were part of a September– November 2018 collection from northeastern and eastern Ethiopian cities Semera and Kebridehar as a part of our previously published studies as previously described [ 6, 43]. Briefly, mosquitoes (n = 7) were collected using Centers for Disease Control and Prevention light traps and pyrethrum spray collection in houses, and larvae and pupae were sampled using the WHO dipping approach. Te mosquito specimens were collected and handled following ethical guidelines as previously described by Balkew et al. in 2020 [ 6], and a materials and data-sharing agreement was established between Baylor University and Jigjiga University. DNA was extracted from the dissected heads and thoraxes of the mosquitos using the Qiagen DNeasy kit. Once the DNA was extracted, the P47Rec ortholog in An. stephensi View in CoL was amplified using two primer pairs. Te first pair (forward—5 ′ -TGGCAAATG ACTAACGTGGA-3 ′, reverse—5 ′ -GTGTTGCCAGTT CGCTGTAA-3 ′) amplified the second and third exons, while the second pair (forward—5-GTGAGCAGCTGT ACGTTGGA-3 ′, reverse—5-AAAACGGAAGGCATG TCATAA-3 ′) amplified the fourth exon. Sequences were aligned using the MUSCLE program and a maximum likelihood tree was generated using the RAxML version 2.0 program [44].