After the chemical or UV mutagenesis, the yeast culture is supposed to be plated on the selective medium with canavanine or 5-FOA in order to assess the efficiency of mutagenesis.
The questions are:
1. after the colonies appear on both plates (regular plate and selective plate) - what is the right formula to calculate the rate ?
2. What should be the normal survival rate after mutagenesis (10/50/90?) in order to get 1 mutation per haploid genome ? (in Molecular Biology Protocols Unit 13.3 , they claim ~60% of survival)
thank you for your comments
Thank you Attila,
It would be even more interesting to find a correlation between the mutation rate and the average number (preferably one per genome) of mutations.. So that to have a meaning rather than just figures (survival / mutation rate)
Hi there,
What you actually measure in your assay is the rate of random mutations abolishing the activity of one specific gene product (CAN1 or URA3). You can't go any further than calculating mutation rate unless you go for global genome analyses.
Thank you Dominique, it is absilutely right. But assuming that the likelyhood of mutation in CAN1 or URA3 is similar to any other gene, we can probably still calculate the average likelihood of one mutation per one genome.
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