Describe 3 ways of measuring changes in mRNA expression and provide an explanation of how each one works. Does the method measure mRNA synthesis, degradation, or levels?© BrainMass Inc. brainmass.com September 22, 2018, 3:02 pm ad1c9bdddf - https://brainmass.com/biology/research-methods-and-experimental-design/mrna-analysis-lab-4886
1. Nuclear run-on assay: Measures relative amounts of mRNA synthesis.
a. Isolate nuclei (keep cold to inhibit transcription) at the different times being examined (e.g., lengths of treatment).
b. Add 32P-labeled rNTPs, allow transcription of nascent RNAs to continue.
c. Hybridize RNA to complementary DNA of interest.
d. Measure level of radioactivity of the DNA-RNA hybrids
An mRNA analysis in the lab are determined. The expert describes three ways of measuring change in mRNA expression. How each one works is provided.