lambda ZAPII clones...
ed at bio02.bio.uottawa.ca
Mon Oct 21 13:01:39 EST 1996
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).
thanks for any help you could give me,
Dep. of Biology
University of Ottawa
PS. incidentally, how do people treat glassware to remove DNA from old
preps ? currently I leave the corex tubes in 3N HCl, but I'm afraid this
isn't nearly enough...any thoughts and wisdom ?
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