lambda ZAPII clones...

Ed ed at bio02.bio.uottawa.ca
Mon Oct 21 13:01:39 EST 1996


Hi all,

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,

Ed Taboada
Dep. of Biology
University of Ottawa
Ottawa, Canada

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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