speI xbaI sublconing

Peter Ellis via methods%40net.bio.net (by pjie2 from cam.ac.uk)
Sat Dec 8 10:01:59 EST 2007


DK wrote:
> aawara from FEMA-trailer.org wrote:
>>
>> Unless the original poster dephosphorylates the ends of his cut
>> vector,
>> it will recircularize very efficiently, and none of his clones will
>> contain the insert.
>
> The situation is no different from blunt cloning. Easy to deal with by
> throwing a large excess of the insert. 20X or more.  With everything
> done properly, a reasonable percent of clones will contain an insert.

Blunt cloning is *also* much easier if you dephosphorylate the vector.

Peter 




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