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Generation of Genetic Cross Parasites
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For performing genetic crosses, WT PfNF54, Pfmacfet¯ and Pfcdpk4¯ were cultured separately to stage V gametocytes, mixed in equal ratio of gametocytemia, and fed to female A. stephensi mosquitoes as described above. Mosquitoes were dissected day 7 post-feeding and midguts were digested to isolate genomic DNA following manufacturer’s instructions from the QIAamp DNA Blood Kit. Transmission of Pfmacfet¯ parasites was determined by genotyping PCRs (Figure 6C).
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