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:


> 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?


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 L. Haviland, PhD
Assistant Professor, Immunology
University of Texas, HSC - Houston
Institute of Molecular Medicine
2121 W. Holcombe Blvd.
Houston, TX  77030

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