Dear all, I want to introduce random mutation to double-stranded DNA phages using the Cobalt60 gamma source. I searched for a suitable dose rate that is high enough to mutate the phages but not too high to damage the phage particle integrity. I found one paper (Bertram 1992) that used 60 Gy/min. Does anyone have any suggestions or documents that I can refer to? Thank you very much.
I think you probably just need to do a dose curve of phage killing. Start with a lysate with known titer and expose at different doses (generally different amounts of time of exposure at a constant distance from the source) and plot PFU remaining after time of exposure. You normally want to be in the range of 10-100 fold reduction of phage titer. If you go much higher then you will start accumulating multiple mutations per genome.
Thank you very much for your answers and information. It is very helpful and informative. Do you have any recommended references that I can look at in more detail?
Thank you very much again. I appreciate it.
Not really, you just need to do some literature searches if want any. There is a quite a lot of older literature on UV mutagenesis and dosage with phages, and I'm familiar with those, but not so much with gamma irradiation. But the principles are the same in either case. However regardless of what you find in the literature, you will need to do a dose response killing curve to standardize your specific equipment and phage. So it might just be easier to do the experiment directly and get the answer since you have to do that regardless.
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