plasmid integration after transfection

Martin Kennedy mkennedy at chmeds.ac.nz
Thu Jan 6 18:41:38 EST 1994


In article <2gglat$h34 at mserv1.dl.ac.uk>, suter at VAX.mpiz-koeln.mpg.d400.de writes:
> question: if one transfects a mammalian cell with a plasmid, how many copies 
> will integrate in the genome ? is this dependend on the procedure followed ?
> 
> cheers,
> clemens

Hi,

The number of plasmids that integrate depends very strongly on the method used
to intrduce the DNA into the cell, for reasons I don't fully understand.  I
know that with electroporation (into mouse ES cells, at least) I generally get
a single copy insertion, and about 10% of the cells have two discrete
insertions.  Very rarely we see more than 2.  With CaPO4 and various other
chemical means, it seems that many more copies integrate, often as concatamers.

Cheers,

Martin

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