Trying to clone DIMERS
Dr. Duncan Clark
duncan at genesys.demon.co.uk
Thu Oct 2 05:25:47 EST 1997
In article <Pine.HPP.3.95.970926190233.8921A-100000 at gong.ci.uv.es>,
Marcos De La Pena Del Rivero <pena at uv.es> writes
> -When the monomers (eluted from a gel) are ligated, I always obtain a
>nice ladder (monomers, dimers, trimers... even hexamers), but the
>eluted dimers cannot be cloned in any vector :(
Could it be that the E.coli you are cloning into just doesn't handle
these types of multimers. Does your PCR product express and could
therefore be lethal with multiple copies? What about trying a lower copy
number vector. I assume you are using a recA(-) (and the rest) host.
The problem with being on the cutting edge is that you occasionally get
sliced from time to time....
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