Help with cotransformation

Michael Banditt banditt at cell.biol.ethz.ch
Fri Feb 16 08:52:43 EST 1996


I am just about to start establishing a cotransformation protocol in 
our lab, in which I want to insert an 8 kb MluI-MluI fragment into the 
genome via homologous recombination together with an episomal plasmid 
carrying a selectable marker. The fragment is hereby cut out of a 
pUC18 derivative by digestion with MluI.

What I would like to know is, whether I have to purify this fragment 
in order to seperate it from the plasmid background before 
transformation, or if this is not necessary. The plasmid itself has no 
homologous sequences to the target gene, except of the MluI-ends of 
course.

Can anybody help me?

--
Michael Banditt                   Phone:  +41-1-633-3345
Institute for cell biology        Fax:    +41-1-633-1069
Swiss Institute of Technology     Email:  banditt at cell.biol.ethz.ch



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