I am trying to make URA3 mutant of an yeast strain. I have prepared FOA plates and when I carry out transformation using salmon sperm dna with my vector containing insert, kanamycin gene flanking URA3 sequences (to delete the URA3 by homologous recombination) , I am getting colonies both in reaction plates and in control (where no vector has been added, only contains wild type strain). The colonies from ligation plates when they are grown in minimal media with uracil are able to grow, but without uracil, there is no growth on plates. How can I check whether the colonies are URA3 mutants or not? I even streaked the wild type on FOA plates, it showed colonies after 36 hours. Is there any biochemical way to test that FOA is working?

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