Ligation, and Transformation iNHIBITORS.

Rich Dudley rdudley+ at pitt.edu
Tue Mar 9 09:04:30 EST 1999


amherrera at CIGB.EDU.CU wrote:

>     Hi all
>
>     I having a low transformation frequency when transform my
>     ligation?
>
>     I have evidence about the good efficiency of the competent
>     cells.
>
>     The vector I used is 10.kb and the band over 3.3kb
>
>     Thanking Any Suggestion
>
>     D.Q.V

How much of your ligation mix are you transforming into what volume of
cells?  I do usually heat my reactions 65oc for 10-15 minutes, then ice
cool and spin down the condensate.  You may have to transform less of
your reaction mix.

Good luck!

rich

--- --- --- -- -- -- --- --- ---
Richard J. Dudley (rdudley+ at pitt.edu)
Research Specialist V
Dept. of Cell Biology and Physiology
University of Pittsburgh
http://www.cbp.pitt.edu
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