I am working on isolation of bacteriophage from soil. My plaque assay looks like this one even after ten-fold dilution of my phage stock. I hope to hear some guide on my work. Thanks!
You don't have a good lawn at all. There are a few possibilities that come to mind:
1) your plating cells are contaminated (or your top agar or buffer is contaminated). I think one of these is most likely the problem.
2) your top agar was too hot and you killed off most of your plating cells.
3) your phage stock is at too high a higher so that even at 10fold dilution you killed most of the plating cells. You may need to make multiple 10-fold dilutions if you have a high titer phage (I would make at 6 x 10-fold dilutions (or 3 x 100-fold) until you know range your phage will be in).
Did you do a control plate with everything other than the phage stock? If you do all the controls you will know where the problem is.
your phage is too high. dillute it a lot and you will see a plaques
- Badges
- Science topic