Disruption of dhfr-ts using a conventional KO construct pBSdh1f8Neo. A) Diagram of the expected genomic loci of dhfr-ts and 1f8Neo in dhfr-ts+/-/Neo parasites. B) PCR analysis with Neo specific primers of WT Tulahuen and both uncloned and selected clones of dhfr-ts+/-/Neo parasites. C) PCR analysis with gDNA from selected clones of dhfr-ts+/-/Neo and WT Tulahuen parasites confirming the expected gene disruption of one allele of the dhfr-ts gene by 1f8Neo. D) Southern Blot analysis of WT Tulahuen and two dhfr-ts+/-/Neo clones digested with SalI and probed with dhfr-ts probe. Diagram not to scale. Numbers are sizes (bp) of expected products.