Plasmid construction

Yulia Shifrin via methods%40net.bio.net (by yulia24 from gmail.com)
Tue May 8 10:19:20 EST 2007


First of all, you can use higher concentration of your plasmid to get higher
yields of these 1bk fragments from the gel. Another option is to amplify
them by PCR.

On 5/8/07, Yvonne Couch <yvonne.couch from dpag.ox.ac.uk> wrote:
>
> Hi all,
>
> I am trying to construct a plasmid and I need to take out some loops from
> one plasmid and insert them into pcDNA3.  The loops are around 1000bp and
> have BglII and BamH1 at either end so they can be digested out.  The main
> problem is the only way I can think of taking the loops out is by
> digesting
> and then running a gel and extracting but this results in tiny amounts of
> DNA that fail to ligate under any conditions.
>
> Any ideas?
>
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