Baculovirus expression system-plaque purification

Keith Johnson kaj at bioc.rice.edu
Mon Oct 14 14:38:22 EST 1996


Hello,

	I am using Baculovirus expression system to purify some protein. Right
now I am trying to purify and obtain a high titer of the recombinant
virus expressing the protein of interest. I am having some problems
isolating the plaques. I used the stain "neutral red" to identify the
plaques but it was not very effective. Then I came across the stain
"MTT" 3-4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (Sigma
product) which has been published to be very effective for the plaque
purification. I am not able to reproduce the staining as described in
the paper even though I am following the protocol exactly as mentioned
(I think!). If anybody has used this stain before or has a better
suggestion for the plaque purification, I would really appreciate their
help. Thanks a lot in advance. My email address is:

mittal at bioc.rice.edu

Thanks

Shalu Mittal



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