PCR Ligation Issues!!!

ChenHA hzhen at freeuk.com
Wed Sep 28 12:06:26 EST 2005


On Wed, 28 Sep 2005 05:58:14 GMT, dk at no.email.thankstospam.net (DK)
wrote:

>
>All kind of things possible but my bet would be with your PCR product 
>not being cut by your enzyme. Blunt clone and excise from the plasmid. 
>BTW, after many grad students in our lab going through similar
>problems, we found that using Invitrogen's  "Zero blunt TOPO" kit
>is a real time and money saver - despite it being really expensive. 
>

I'm sure it works well, but I do wonder whether these kits help with
the research work or just let people who don't know what they doing to
get by.  I personally would not use something like TOPO cloning kit,
primarily because the cloning method I use works well (haven't any
cloning problem yet after many years), and also because it is
expensive and not really easier for my purposes.  Although if I do
come across something really difficult to clone I might consider using
it.


>DK



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