sindbis plaque assays??

Brett Lindenbach brett at BORCIM.WUSTL.EDU
Mon Mar 8 16:39:39 EST 1999


>I am looking for some hints on plaque assays for sindbis virus.
> Do you have to use chicken embryo fibroblast cells for this?  Do you use the
>agarose overlay method or can you just stain with crystal violet?
>
>Thank you for any information....
>
>Bolyn


Depends on what you want to do.  If you are using a SINrep vector or
studying SIN biology and are titering your stocks, you should titer the
virus in the cells you will do your experiments in.  If you just want to
quantitate virus, say from an infection, you do not have to use CEFs.
BHK-21 cells work well, although yield around 2x less plaquing efficiency
than CEF cells.  You will need to use an overlay in either case since SIN
will spread pretty quickly through a culture.  I'd allow your plaques to
develop for around 2 days then fix and stain with CV.

Brett Lindenbach
brett at borcim.wustl.edu
Dept of Molecular Microbiology
Washington University School of Medicine
(314)362-2767





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