Controls in stable transfection experiments

David L. Haviland, PhD dhavilan at IMM2.IMM.UTH.TMC.EDU
Sun Feb 28 11:13:34 EST 1999


On 28 Feb 1999, Wolfgang Schechinger wrote:

*EDITED*

> Now what should the appropriate control cell line be like?
> 
> - transfected with empty vector (expresses only the neomycine 
> phosphotransferase)
> 
> - transfected with another non-related protein (would be under the 
> same stress of protein overexpression)
> 
> - untransefected
> 
> The point is: If I transfect the empty vector, do I really get a 
> control? I get the NeoR expressed, but since the vector is inserted 
> *somewhere* in the genome, will I really produce a control cell line?
> 
> What's generally accepted?

Wolfgang:

There isn't much of a "convention" here as to what to use.  But for the
record, other investigators in my group have used an empty vector as a
control and the review editors didn't argue.  Of recent, I've seen some in
the literature using their "favorite gene" in the same vector but in a
*reverse* orientation as a control for their gene being in the correct, or
forward, orientation.  

Hope this helps,
David

==========================
David L. Haviland, PhD
Assistant Professor, Immunology
University of Texas, HSC - Houston
Institute of Molecular Medicine
2121 W. Holcombe Blvd.
Houston, TX  77030
http://www.uth.tmc.edu/~dhavilan
713.500.2413-Voice
713.500.2424-FAX
==========================






More information about the Methods mailing list