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Best way to express Lucif activity?

Joe Miano jmiano at post.its.mcw.edu
Tue Oct 8 22:02:56 EST 1996


Thanks for the note.  I have always wondered with suspicion about
contransfections.  Does this not introduce squelching and perhaps an
under-representation of the true promoter activity? I have seen some labs
not use an internal control plasmid but I think you're right if you say a
majority of labs do.

What I typically do is transfections in triplicate (6 well dishes) and I do
this at least 3 times.  I am finding no significant difference between
experiments when I express the normalized RLUs (normailzed to protein) as a
percent of SV40 or as a fold increase over pGL3basic (which bythe way gives
high background in many cells!).  I do not see any profound argument
against not using an interbnal standard although I can imagine some hostile
reviewers wanting one to do it the "correct way."  

Is there really a correct way of expressing the data?

I would encourage more people to comment on this issue that I have debated
for some time know.  I really do not know the answer.

The Love you take is equal to the Love you make--P. McCartney

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