ONT of T. gondii ME49 brain cysts
DNA was extracted from the collected cysts. Genomic DNA waspurified using a phenol-chloroform extraction protocol. Briefly, T. gondii cells (5  10^7)were washed in PBS and resuspended in a digestive solution containing 500 l TNE, 50 l10% SDS, and 7.5 l Proteinase K (10 mg/ml). The suspension was incubated overnight at56C with shaking. Following lysis, 500 l of chloroform/isoamyl alcohol (24:1) was added,mixed vigorously, and centrifuged at 13,000 rpm for 5 minutes at room temperature. Thesupernatant was transferred to a new tube, and the extraction was repeated. DNA wasprecipitated with 500 l isopropanol, incubated for 5 minutes at room temperature, and centrifuged at 8,000 rpm for 10 minutes. The DNA pellet was washed three times with 500l of 70% ethanol, dried for 10 minutes at room temperature, and resuspended in 100 l TEbuffer (incubated for 1 hour at 56C, then overnight at 4C).