Kanamycin selection

Richard J. Dudley rdudley at nrc.uab.edu
Wed Feb 25 12:56:42 EST 1998


Jun-Ichi Aikawa wrote:
> 
> What concentration of kanamycin is used for the section of recombinant E.
> coli? I bothered high background of non-recombinant clones. My conditions
> are as follows:
> kanamycin: 30 ug/ml in LB plate (according to Current Protocol)
> E. coli: XL2-Blue MRF, XL1-Blue, DH5a (commercial competent cell)
> plasmid: pBK-CMV, pEGFP-N1
> 
> Please give me any suggestion and comments.
> 
> Jun-ichi Aikawa, Ph.D.
> University of California, San Diego


Are you sure your kanamycin is 30 ug/ml?  Check the potency of your
batch of kanamycin.  The lot I just bought from Sigma is 784 ug kan/mg
of powder in the bottle.  You'll have to adjust the mass you add to get
a true kanamycin concentration of 30 ug/ml.

rich
-- 

--- --- --- -- -- -- --- --- ---
* Yes, Kathy really did let me name the cat Histidine. *

Richard J. Dudley (rdudley at nrc.uab.edu)                            
Department of Neurobiology                    
University of Alabama School of Medicine       
http://www.nrc.uab.edu/



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