Multiple plasmid FuGENE 6 transfections
Ian A. York
iayork at panix.com
Sat Sep 22 08:48:01 EST 2001
In article <3BACF420.16055.1988A5 at localhost>,
Wolfgang Schechinger <wolfsc at ibms.sinica.edu.tw> wrote:
>In contrary, I would like to be sure that only one copy enters a cell (I
>testing a library on eucaryotic cells). What methods for transfection would
Electroporation *tends* to put fewer plasmids per cell, but I htink there
are still many.
There's a protocol in which you fuse the membranes of your
plasmid-containing bacteria with your cells for transfection, and then
dump on a ton of antibiotic. That's supposed to put a single plasmid per
cell. (Actually, of course, many plasmids, but all identical clones.) I
believe it's in the Red Book of molecular biology protocols. I've never
tried it myself.
>What about multiple stable transfection of individual cells using retrovirus (I
>currently do that and assume ~one infection per cell)
How many selection markers do you have?
If you're testing a library, are you planning on re-cloning your library
into the retrovirus?
Ian York (iayork at panix.com) <http://www.panix.com/~iayork/>
"-but as he was a York, I am rather inclined to suppose him a
very respectable Man." -Jane Austen, The History of England
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