Transfection of Sf9/Sf21 Cells

Elmer M. Price, Ph.D. DCRCEP at mizzou1.missouri.edu
Wed Jun 8 09:45:45 EST 1994


In article <garry.23.0011B4AB at menzies.su.edu.au>
garry at menzies.su.edu.au (Garry Myers) writes:
 
>
>Hi,
>
>I've recently become the proud owner of the CLONTECH BacPAK expression
>system ;).  The protocol accompanying the kit suggests using Lipofectin(tm)
>to transfect the Sf21 cells with recombinant virus.  I'm interested to hear
>any thoughts or opinions from baculoviral-minded individuals on other
>methods of transfecting Sf9/Sf21 cells, particularly electroporation.
>
>Cheers,
>
>Garry.
>
>___________________________________________________________________
>Garry Myers                       Menzies School of Health Research
>Molecular Genetics Unit           garry at menzies.su.edu.au
>___________________________________________________________________
Hi Garry-I would strongly recommend against electroporation as a means
of transfecting insect cells.  The point of the transfection is to
introduce two DNA molecules into the insect cell-one being the transfer
vector and the other being the 140Kb baculoviral DNA. Once inside the cell,
the miracle of homologous recombination occurs and recombinant baculovirus
is generated.  This is not too terribly efficient and in order to optimize
the entire process, one needs to optimize the co-transfection process.  The
lipofection process works very well, while electroporation is probably (IMHO)
not as efficient at co-transfection (but great for single transfections).
The lipofection step is also very easy, taking only about 30 minutes of effort
on the part of the investigator.
 
Hope this helps
 
Elmer Price, Ph.D.
 
 



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