Fig. 3.

BIR product formation is delayed for the 70-kb donor strain compared with the 15-kb donor strain and occurs with highest efficiency in G₁ released cells. (A) FACS profiles and PCR to detect DSB (D primers) and BIR product (P primers) from cells of the indicated strains that were arrested in G₁ and released at the time of HO induction. (B) Kinetics of BIR product formation in G₁-released cells determined by the ratio of BIR to control (C) PCR products from three trials; error bars show the mean ± SD. (C) FACS profiles and PCR to detect DSB and BIR product from cells that were arrested in G₂-M at the time of HO induction. (D) Kinetics of BIR product formation in G₂-M arrested cells; error bars show the mean from three trials ± SD. (E) Schematic showing sizes of digestion products for the recipient chromosome before and after HO cutting, the donor chromosome and BIR product. The pale orange bar indicates the position of the hybridization probe and the vertical arrows show the location of EcoRV sites. (F) Southern blot of EcoRV-digested genomic DNA of the indicated strains before and after HO induction.