Skip to main content
. 2010 Jul 8;6(7):e1001007. doi: 10.1371/journal.pgen.1001007

Figure 5. Gene conversion assay.

Figure 5

(A) Schematic of the gene conversion assay. I-SceI cleaves the plasmid to generate a linear fragment with homology to the ADE2 locus at both ends of the break. Repair from the genomic locus results in restoration of the AatII site, replacing the I-SceI site. The location of PCR primers used to identify extension of the invading strand strand and completed products are shown by arrows labeled e, f, g and h. (B) Southern blot of genomic DNA digested with BamHI (B) to liberate a 3.7 kb fragment containing the plasmid ade2 allele. Cleavage by I-SceI results in fragments of 2.0 and 1.7 kb and repair restores the 3.7 kb fragment. (C) PCR to detect initiation of DNA synthesis from the left invading end at different times after induction of I-SceI. (D) PCR with primers g and primer h to amplify the plasmid ade2 allele followed by AatII digestion to detect repair of the I-SceI site.