FIG. 6.

Real-time gene conversion assay. (A) DNA from MK301 and MK301rad24 cells was extracted at intervals after HO induction. Equal amounts of PCR products flanking the DSB were digested with BamHI. In this strain, homology length is 12.8 kb, and therefore the donor URA3 sequences are also detected in the PCR assay. At 0 h, only half the PCR products can be digested with BamHI. Repair of the broken chromosome by gene conversion progressively increases the relative proportion of BamHI-containing fragments. Uncut PCR fragments represent the portion of the population that has not undergone gene conversion. (B) Slot blot assays. Nondenatured DNA was hybridized with probe B (complementary to 1.2-kb URA3, as described previously). (C) Graphic representation of the kinetics of gene conversion. Percent gene conversion represents the relative percentage of PCR fragments digested by BamHI compared to those seen at 0 h.