This PDF file includes:

• fig. S1. The cell ploidy and the choice of reference mRNAs have no impact on measured half-lives.
• fig. S2. Pop-out of the chromosomally integrated plasmid is dispensable for half-life estimation of mature mRNA.
• fig. S3. The doubling time of the MGC strains (red dots) is similar to that of the parent diploid strain (black line).
• fig. S4. Insertional promoter strategy.
• fig. S5. Phenanthroline increases the number of P bodies.
• fig. S6. mRNA half-life has a major impact on response time of a protein in a negative feedback loop.
• fig. S7. Degradation rates and not synthesis rates affect the response time.
• fig. S8. Parameter estimation of the two-state model to fit the USA1 RNA copy number distributions.
• fig. S9. Impact of 5′UTR length on the decay profiles of mRNAs with varying 5′UTR lengths.
• fig. S10. Design of the plasmids with the substitutional promoters and the strategy for chromosomal integration.
• fig. S11. Reverse transcription strategy.
• table S1. Plasmid list.
• table S2. Strain list.
• table S3. Cloned 5′UTR lengths of genes.
• table S4. qPCR primers detecting marked mRNAs and their amplification efficiencies.
• table S5. qPCR primers detecting endogenous mRNAs with their efficiencies.
• table S6. Cross-reaction of primers to detect the marked mRNA with the endogenous mRNAs.
• table S7. Mean, Fano factor, and CV of the RNA molecule copy number distributions.

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Other Supplementary Material for this manuscript includes the following:

• data file S1 (Microsoft Excel format). Ct values for one of the replicate experiments of pooled strains in the MGC experiment.
• data file S2 (Microsoft Excel format). Time courses in the replicate measurements.
• data file S3 (Microsoft Excel format). mRNA (relative) levels and decay rates.
• data file S4 (Microsoft Excel format). Time courses for YJR139C and YDR032C mRNAs.

Download data files S1 to S4