Genome of Australian fly defeats killer bacteria by evolving to co-exist with it PLOSBiology
overlapped . Instead, we initially designed reverse primers to distinguish the 2 strains via standard PCR, based on alignments from theMLST sequences we generated for the female-biased and non-female–biased lines and used these in conjunction with the existingforward primers to produce bands of 373 bp and 388 bp that amplified the female-biased and non-female–biased strains, respectively. The PCR conditions for both primers were the same .
A subset of the isofemale lines initiated from field collections including 98 lines from Brisbane, 11 from far northern Queensland, and 2 from Nowra were screened with these primers. To determine relative densities of the strains, we later used genomic sequences to design strain-specific primers specifically for use on the Roche LightCycler 480 system .
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