wind at biobase.dk
Thu Jun 6 03:26:25 EST 1996
One way of obtaining pure phage particles is to do a CsCl-gradient
purification. You may have to dialyse afterwards to get rid of the
salt, but IMHO its worth the effort.
Let me know if you want a protocol, I'll try to dig it up.
DANIEL Y KIM (dkim at nmsu.edu) wrote:
: I have been trying to construct a filamentous phage display of a protein
: fused to M13 gene III. In my plasmid/phagemid, the gene III fusion is
: interrupted by an Amber stop codon. I imagine there is a fair amount of
: protein being produced which is not fused to gene III and which will be
: secreted into the growth medium, instead of being incorporated into a
: phage particle.
: Is there a way to precipitate the phage without the free protein? It
: seems that PEG will precipitate both phage and free protein (60 kDa) out
: of the medium (positive Western signal in the absence of helper phage).
: If not, is there a convenient way to filter out the larger phage from the
: free protein?
: Daniel Kim
: dkim at nmsu.edu
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