Sonication of Vaccinia virus

Koen Allosery koen_allosery at innogenetics.be
Mon Sep 13 10:52:40 EST 1999


Geoffrey Cole wrote in an article in 'bionet.immunology' :

> ...
> Does failure to sonicate fresh thawed aliquots of virus (infected cell
> lysates) significantly reduce available infectious virus?  Is placement
> of a thawed vial of virus in an open bath sonicator for 1-2 minutes
> sufficient to break up virus aggregates.
> ...

Same discussion in our lab !

The argumentation in our lab to sonicate the Vaccinia virus stocks prior to
each use, was the presence of cells and cell debris in the lysates.  The
virus would stick to this components and form aggregates by doing so.

Since we clear the harvested virus (after 3 freeze-thaw cycles) by
centrifugation (10 min @ 230g / 4°C to remove cells and 10 min @ 1350g /
4°C to remove cell debris), this should not longer be necessary.  The virus
is stored in serum-free medium.

However, we never tested the difference in titres between sonicated and
non-sonicated virus stocks and kept sonicating our 1.5 ml-vials for 2 min
in an open bath sonicator.

Our internal debate here is still open until one of those days we will
perform a plaque assay to end the discussion.

Any comments ?




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