Hello,
I tried to stain some phages with SYBR green I (Roth) to do some epigenetic analysis. The phages were from a commercial source, and I stained 500 uL of it with 5 uL SYBR green for 15 min in the dark.
In the attached picture, one can notice (1) very few phages (I was expecting thousands of dots, since the given concentration of the preparation is about 10^9 PFU/mL); (2) a possible phage (yellow arrow) and (3) some garbage or a phage aggregate (red arrow).
Is there a good technical protocol for epifluorescence of phages? What controls should I use to confirm the presence of phages in the slides?
Thank you
not sure that what you saw were phages: with epifluorescence you shoul only see a milky way of phages if they are well individualized: I think that what you saw are aggrates or cells debris. furthermore I doubt that SYBR green could enter the phage capsids that are supposed to be very tough with a very dense DNA in them...
I agree, but the protocol I found only says to stain the preparation at room temperature for 15 min. Is there a better one?
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