i am trying to purify bacteriohage phix174, in gradient i got one separate band. but when i see that sample in negative stain, i got some kind of contamination in sample either bacterial membrane or DNA.
any body have experience in purification of phage and EM analysis that what kind of contamination i have in my sample.
Hello Saleem,
In my opinion, you need to use freshly amplified phage particles for performing TEM. Usually, people have purified virus by CsCl density gradient using general centrifugation. You may use 0.75% Uranyl formate for the negative staining. If you need further information, you may follow this article instructions. I hope this information may help you.
Article Virus purification by CsCl density gradient using general ce...
i think this is a degradation problem of phage, i purified phage but become unstable after 24 hours
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