cloning the wrong way?

Jens Tornoe jens at tornoe.net
Wed Feb 16 15:53:51 EST 2000


Apparently, your insert seems to be toxic to the cells. I think Stratagene
has a kit for cloning such difficult genes. It could be using the SURE
cells, but I am not sure (no pun intended).

> Do I have to sequence the clones to make sure that they are the right
> DNA and to confirm the orientation?

You don't need to sequence your clones to confirm your orientation (your
digest did that) but it is always a good idea to sequence a few clones when
cloning PCR products. This is especially important when you use Taq
polymerase, which has an higher error rate than e.g. Pfu.

>
> Any comments are greatly appreciated. Please email me privately as well
> to my email address.
>
> Cheers,
>
> Ahmed
>
>
> Sent via Deja.com http://www.deja.com/
> Before you buy.






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