Kanamycin selection

Sebastian Bunka bunka at i112pc09.vu-wien.ac.at
Thu Feb 26 06:32:46 EST 1998


Jun-Ichi Aikawa <jaikawa at UCSD.EDU> 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

Sambrook 2nd. ed.: stringend plasmids 10ug/ml, relaxed plasmids 50ug/ml.
We use generally 50ug/ml. Works for us. But check the label on
your Km vial. Mostly Km is supplied not pure or at 100% potency!!

For ex. Sigma Kanamycin is labelled as 75% potency, means if you weigh
30ug/ml you will only have 22.5ug/ml active Km!

SWB


-- 
Sebastian Bunka, Dr. med. vet
Inst. Med. Chemistry, Vet. University Vienna
Ph. +43-1-250 77 ext. 4208, Fax: ext. 4290
e-mail: Sebastian.Bunka at vu-wien.ac.at



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