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How to publish real time PCR results?


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#1 Nephrite

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Posted 22 November 2018 - 02:46 AM

Dear all,

I work on analyzing the expressions of some enzymes in muscle tissue, myoblast and myotube cultures. The aim of the study is to demonstrate presence or absence of expression in my samples.

I isolate RNA, I measure it, I check the quality on gel, I generate cDNA, I measure it, I do the qPCR. The melting curve analysis shows single products, which can be seen on the gels. I have done enough biological and technical repeats. Unfortunately my cycler can run simultaneously only 36 samples, which makes the job very slow. The targets are 9, and they are normalized versus 2 refferent genes. Anyway, the results repeatedly show that the cultures, unlikely to the tissue, do not express some of the enzymes.

The question is how to publish this? I read some MIQE guidlines, they require calibration curve for each target. Should I really do this and how? Should I include in the paper the melting curves for each product? What proof would be enough to show precision of the experiments?



#2 bob1

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Posted 26 November 2018 - 08:53 AM

As you are just going for presence/absence, you shouldn't have to perform any analysis of the data, indeed it shouldn't matter whether it was done on a qPCR machine or not. I would just run the products on gels and add that as a supplementary image.



#3 Nephrite

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Posted Today, 01:24 AM

Thank you, Bob1! :-)






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