Ed wrote:
> I had a little question...I have screened a cDNA library made in Lambda
> ZAPII, which has self excision of pBluescript when the phage is plated with
> a certain helper phage (which we conveniently do NOT have >:( ). Now, my
> question is...since budgetary constraints are a bit of a problem (and
> getting the kit for excision is a huge waste, since I'll be using it once !)
> should there not be a way of cutting out the pBluescript out of the
> lambda phage, and re-circularizing it ? I mean...from the vector map, it
> looks as though the whole pBluescript vector is there, and the automatic
> excision is nice, but not necessary to get your inserts out into pBluescript
> without too much trouble...I think all I would need would be to cut out
> the bluescript (and the insert within it) and re-circularize it
> ...has anyone ever done this before ? It just seems silly to cut the
> insert out and clone into bluescript, when it is already in bluescript
> (although in a linearized form).
A quick glance at the Stratagene catalog informs me that the ExAssist phage (which
I've used with excellent results) are $95.00. Not much when you consider the price
of many restriction enzymes. And the phage are stable for years in the freezer
(the tube I used was over 2 years old and had been purchased by an earlier
post-doc), so who's to say you'll only be using it once?
----------------
John Watson
Bristol-Myers Squibb Co.
watson_j at bms.com
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"If you're not part of the solution, you're part of the precipitate."
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