4 way ligations?

vilimf01 at mcrcr6.med.nyu.edu vilimf01 at mcrcr6.med.nyu.edu
Sun Jun 23 10:23:04 EST 1996


A reason wich immediately came to my mind is that the oligo's are not
phosphorylated. But you probably thought of this already.  The second
thing to consider is that if all the oligo's are kinased, you could
get multimerization of your linkers, depending on the exact
configuration.  Best thing to do is to treat them as linkers and
ligate them onto the insert first (you can avoid multimerization by
appropriate kinasing of the linker oligo's). Just my 2 cents, hope its
of some use.
					Regards		Sven


In article <DtDCtJ.2pI at news.hawaii.edu>, angel at ahi.pbrc.hawaii.edu (Angel Yanagihara) writes:
> Hello...
> 
> I'm attempting a 4 way ligation and was wondering if anyone out
> there has had any luck with this kind of thing.  I have built two
> overlapping oligo stretches (about 60bps each) to link my insert
> (about .8Kb) by bridging the 5' side of my insert to one side
> of my vector and the other overlapping oligo is designed to
> bridge the 3' side of my insert to the other open end of my
> vector.  All sites involved are sticky end, unique, and compatible
> only with each unique hookup involved.  I've tried to ligate
> them all together in one ligation reaction, as well as attempting
> various kinds of sequential ligations, with no success.
> 
> Any suggestions with this would be greatly appreciated
> 
> aloha and mahalo
> Angel
> 
> angel at halia.pbrc.hawaii.edu
> 



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