single colony PCR

Godfrey Godfrey.Neutelings at univ-lille1.fr
Tue Jan 25 10:37:29 EST 2000


dans l'article v04210101b4a54a3574ef@[128.206.80.4], "Peter L. Schuerman" à
schuermanp at missouri.edu a écrit le 14/01/00 23:00 :

>  Can anyone recommend a good method for single-colony PCR that is more
>  elaborate than simply using cells directly in the reaction?  In
>  particular, I'd be interested in a method that works for
>  Agrobacterium.  In my experience directly adding cells works well for
>  E.coli but I have been getting poor amplification and inconsistent
>  results when I have tried this with Agrobacterium.  Any help would be
>  greatly appreciated.
>
>  Regards,
>  Peter L. Schuerman


Simply pick a single colony and place it in 50 microL of water. Boil for 2
min, place on ice. 5 microL is enough for a 50 microL PCR reaction.
Works fine

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