Subcloning pcr products

David Michael Eisenmann eisenman at leland.Stanford.EDU
Fri Oct 13 19:27:49 EST 1995


imc1001 at cus.cam.ac.uk (Dr I.M. Clark) writes:

>We have produced an approx 500bp pcr product with a SalI site at 
>one end and a BamHI site at the other.  We have tried to cut and
>subclone this into a Sal/Bam cut vector with no success.
>--

>Rheumatology Research Unit,
In the back of the NEB catalog is a table of how well different enzymes
cut depending on how many bases are left from the end of the fragment.
One of your enzymes may be cutting poorly.  I seem to recall having trouble
with Bam myself in the past.  As others have suggested, we now almost
always subclone PCR products into the Invitrogen TA cloning vector
and then clone them out of that.

Dave Eisenmann
Stanford University
>Addenbrooke's Hospital,
>Cambridge. UK.



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