An easy tool to monitor the elemental steps of in vitro translation via gel electrophoresis of fluorescently labeled small peptides

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FIGURE 5.
FIGURE 5.

Monitoring translation termination efficiency by urea PAGE. MF-coding mRNAs with different stop codons (lanes 1, 4—UAA; 2, 5—UAG; 3, 6—UGA) were translated in vitro in the presence of BPY-M-tRNAfMet by PURExpress ΔRF123 Kit with the addition of either RF1 and RF3 (1–3) or RF2 and RF3 (4–6) RFs as indicated above the lanes. Lane 7 corresponds to the BPY-M-tRNAfMet. (A) Separation of translation products without chemical post-treatment. (B) Separation of translation products after hydrolysis by 1 M NaHCO3. BPY is marked with a green oval—BODIPY label; black circle—methionine; gray circle—phenylalanine residue; blue oval—tRNAfMet.

This Article

  1. Published in Advance December 26, 2023, doi: 10.1261/rna.079766.123 RNA 30: 298-307

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