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. 2016 May;22(5):657–659. doi: 10.1261/rna.056325.116

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© 2016 Garcia and Parker; Published by Cold Spring Harbor Laboratory Press for the RNA Society

This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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FIGURE 2. — Decay fragments are detectable in MFA2-MS2 genomic constructs. Total RNA was isolated and visualized from strains expressing MFA2-12×MS2 from the chromosome. Grown as described in Haimovich et al. (2016). (A) Total RNA from wild-type, mfa2::MFA2-12×MS2 (MFA2-MS2) and mfa2::GAL_promoter-MFA2-12×MS2 (gGAL-MFA2-MS2) plus and minus the pUG36-CP-GFP×3 plasmid was run on a 1.5% formaldehyde agarose gel, probed with 12×MS2 probe, and then stripped and reprobed with 7S probe (5′-GTCTAGCCGCGAGGAAGG-3′). (B) Same as above except visualized with a probe to the 3′ UTR of MFA2 (5′-GATGAGAGAATTGGAATAAATTAGTTTGCCAGC-3′) with a light (top panel) and a dark exposure (bottom panel) of the blot. Quantification of the percentage of decay fragment relative to full-length MS2-tagged mRNA was done using ImageJ to calculate the total intensity of the decay fragments over the total intensity of the full-length mRNA. Both values had the background values subtracted prior to calculation. This percentage represents the average of two biological replicates and its standard deviation.