Figure 2.

Gel electrophoresis images depicting the process for identification of mutant numbers 99–122. A total of six individual mutant DNA samples of the 25 that had been pooled for sequencing, were pooled, and subjected to amplification using primers designed for the mutant of interest (Fig. 2A). Individual DNA samples for reactions that yielded a product of the expected size were separately amplified with the same primers (Fig. 2B). To verify the correct insertion locus, primers that amplified across the entire insert and flanking regions were used (Fig. 2C). Stars in panels A, B indicate the isolate with the mutation of interest. Primer combinations are indicated above the black bars, and mutant isolates are listed below the black bars. In panel C, mutant 100–1 to 100–8 and 116–1 to 116–8 denotes populations of that mutant isolate obtained by limiting dilution. The presence of a single high molecular weight amplification product indicates that there are no wild-type or other mutant A. phagocytophilum in these populations.