To isolate Neurospora haploid auxotrophs from prototrophs yo

To isolate Neurospora (haploid) auxotrophs from prototrophs, you perform a large-scale mutagenesis. You learned in class that you should culture them on the complete medium first and then test them on the minimal media plate by making replica plates. But, there is a way to do it using the minimal media plate ONLY. Dr. T laid out a basic plan below. He left out culture conditions of Neurospora for this experiment. Complete the plan by filling in the boxes and indicate which ones are auxotrophs (use dots wisely). Note that we usually culture Neurospora at 30degreeC. You are not allowed to add any amino acid supplements to minimal media plates. In other words, you can use only minimal media plates in this question. Think about environmental conditions other than amino acid supplements.

Solution

1. The spores are mutagenized using X-rays, then these mutagenized spores are grown on complete medium (minimal medium plus a full range of aminoacids).

2. The minimal media is supplemented with one amino acid. Spores from each culture grow on one and only one of the supplemented media.