This assay was performed as previously described (11). Cells were grown in YP media containing 2% raffinose to an OD600 = 1.0 at 30°C. To induce HO endonuclease expression, cell cultures were adjusted to OD600 = 0.5 with YP media containing 2% raffinose, and 2% galactose was directly added. Genomic DNA of each specimen was extracted every 1 hour. PCR analysis of genomic DNA was performed using the following primers: for the detection of DSB generation, 5′-CTTTTAGTTTCAGCTTTCCG-3′ (pI)/5′-ACTCTATAAGGCCAAATGTACAAAC-3′ (pJ); for the detection of strand invasion and primer extension process, 5′-GCAGCACGGAATATGGGACT-3′ (pG)/5′-ATGTGAACCGCATGGGCAGT-3′ (pH); for the detection of ligation process, 5′-AGATGAGTTTAAATCCAGCATACTAG-3′ (pC)/5′-TGTTGTCTCACTATCTTGCCAATAAG-3′ (pD); for the detection of ARG5, 6 as a control DNA region, 5′-CAAGGATCCAGCAAAGTTGGGTGAAGTATGGTA-3′/5′-GAAGGATCCAAATTTGTCTAGTGTGGGAACG-3′.

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