cloning problem

Kyle Legate legatek at mcmail.cis.mcmaster.ca
Sun Nov 17 11:12:25 EST 2002


On 17 Nov 2002 nazarian80 at yahoo.com wrote:

> hi
> I have attempted to clone a 1.3 kb insert (Bamh1/Xho I) into a PRSETA and PET28a vector (Bamh1/xho1)
>   The vector has been gel purified and  fragments ligated (NEB T4 ligase overnight at 14 C),
> and transformed into BL21 DE3 cells .   After
> plating/overnight incubation, very few colonies are present OR not found, and after screening
> for positives with Bamh1/Xho I, there is no insert.  Any suggestion/tips?
>
Try again with different cells. BL21 DE3 are an overexpression strain, not
a cloning strain.

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legatek at mcmaster.ca		Kyle Legate            legatek at hotmail.com

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