Help!!!Restriction enzyme digestion troubles.

ChenHA hzhen at freeuk.com
Tue Jun 27 21:45:50 EST 2006


On Mon, 26 Jun 2006 08:03:45 +0100 (BST), shyam nandi
<biotech_iicb at yahoo.co.in> wrote:

>i am doing  double digest restriction digestion of baculogold vector with
>  RE pstI &  kpnI
>  the both sites are contigously present one after another in the vector
>  liki 
>                            .....CTGCAGGGTACC......      (vector)
>                                 pstI        I       kpnI
>   
>  Is my digestion should complete successfully.
>   

Probably not.  Digest with PstI first, then KpnI.   

Check NEB catalog for information on how many bases are required for
good digest.



>  Please reply
>   
>  i am not getting any colony after ligation & transformation
>   
>  While getting colony in non contiguously present  RE site,using same vector
>   
>




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