IUBio

Conjugation?

Edgar Valencia edgar at CIFN.UNAM.MX
Wed Mar 3 18:05:48 EST 1999


Hi Mounir
I work with Rhizobium and usually for biparental matings I use a ratio 2:1
Recipient-donor, in a 150 microliters "reaction" For triparental
matings I use 80 microliters of the receptor, 40 of the donor and 30
of the helper which gives you an almost "1 : 1" donors recipient. What
I also know from experience is that ampicillin
DOES NOT WORK in our Rhizobium as a marker. The plasmid gets in, but you
cant select for ampicilin resistant colonies, you have to use another
marker in the plasmid. I work with R. etli, perhaps your Rhizobium behaves
different, but I suggest you to keep your eyes open 
Good luck
 ###################################################################
 # ______ #
 # \ \____ # 
 # Edgar Valencia-Morales ********]|-----|___\____ #
 # Departamento de Genetica Molecular ********]|_______>______/ #
 # CIFN - UNAM /______ / #
 # MEXICO #
 # e-mail edgar at cifn.unam.mx #
 # #
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