Convenient and Acceptable Versus Appropriate and Ethical - Convenient and Acceptable Versus Appropriate and Ethical (Nov/18/2008 )
In our lab we had an informal discussion on what or which kind of results are appropriate to consider and many a times which we take it as genuine because some other lab does it or if journals accept it. Just for example, :
1. Can we compare two gels of PCR or Western blot performed and ran on two different days?
2. Can loading control be ran on another gel and used for our test samples?
3. Can internal differences can be ignored when comparing different normal samples from human patients?
4. Which is the best normalization control?(GAPDH, RPL, B-Actin and many others may differ for example between normal and cancer)
There are so many other such queries that arise but how to decide on these? Should we follow some unwritten rules just because they are convenient for us and acceptable for journals or should we dig deeper in to this so that we get a genuine and appropriate results and observations! I am really glad that my PI always insists on appropriate controls and repeats.
Calvin*
True! Those ain't science!
1. Can we compare two gels of PCR or Western blot performed and ran on two different days?
Quantitatively??? People do that?
I mean, the whole point of comparing gels from different days is to repeat an experiment and see if the results also repeat, or you mean something else...
Those ain't science??! I have to disagree here.
1. Can we compare two gels of PCR or Western blot performed and ran on two different days?
Yes, we can. Why couldn't we? If the gels are done with the appropriate controls, they certainly can. Of course, you can't quantify from one gel to another, but for qualitative purposes, sure.
2. Can loading control be ran on another gel and used for our test samples?
Same thing. Do not forger that Westerns are a semi quantitative evaluation of protein presence.
3. Can internal differences can be ignored when comparing different normal samples from human patients?
No. Every parameter should be taken into consideration.
4. Which is the best normalization control?(GAPDH, RPL, B-Actin and many others may differ for example between normal and cancer)
Each normalization control should be appropriate to the specific experiment you do. Actin may not be the best control for exp A, while it is for exp B.