TA cloning vector

Stuart Brown browns at ccu.umanitoba.ca
Fri Aug 6 14:47:37 EST 1993


>Anybody have an opinion as to whether those tenacious enzymes would still
>be stuck to the 3' end after an hour at 100V in 2% NuSeive?  I ask in all
>seriousness, as this PK step is mentioned relatively often, and I want to
>start cloning PCR products excised from agarose gels in the near future.
>
>Alex Parker
>U Maine Zoology
>

I have no problem cloning PCR products after running in a gel, cutting
out the band, and extracting DNA by gene clean (glassmilk proccedure)
and then using the Invitrogen TA kit.  I have also had success with the
same type of glassmilk extracted DNA cloned by restrction digestion,
cleaned by phenol extraction and EtOH ppt. 

Gel electrophoresis is the best way I know of to remove proteins from
DNA.

	-Stu
	
-- 
Stuart M. Brown                             If you can remain cool when all 
U. of Manitoba, Dept. Plant Science         Around you are in panic,
Winnipeg, Manitoba, CANADA
browns at ccu.umanitoba.ca            Then you surely misunderstand the situation



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