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

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     #                                             ______              #
     #                                             \     \____         # 
     #       Edgar Valencia-Morales        ********]|-----|___\____    #
     #  Departamento de Genetica Molecular ********]|_______>______/   #
     #           CIFN - UNAM                        /______ /          #
     #             MEXICO                                              #
     #     e-mail edgar at cifn.unam.mx                                   #
     #                                                                 #
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