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Reverse transcription
equal mass
equal volume
mRNA Stability
Mouse Pluripotent Stem Cells

Why use equal mass instead of equal volume for reverse transcription?

sy
sheng yang Mar 4, 2025
protocol Protocol: mRNA Stability Assay Using Transcription Inhibition by Actinomycin D in Mouse Pluripotent Stem Cells

I hope this email finds you well. I apologize for the interruption, but I have a question regarding RNA normalization in reverse transcription.

Why is equal mass of RNA used instead of equal volume? As treatment time progresses, the total RNA extracted from the same number Read more Read less more less

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Minna-Liisa Änkö Author Answered Mar 18, 2025

Centre for Reproductive Health and Centre for Cancer Research, Hudson Institute for Medical Research, Australia

We considered both options (mass and volume) but the volume is very prone to fluctuations. Due to the relatively short treatmetn time, the equal mass was still an appropriate way of normalisation. This had also been used in the literature by others.

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