FIGURE 3.
Roadblock-qPCR yields accurate measurements of mRNA half-life. (A) Reporter mRNA stability analysis using transcription shutoff with doxycycline. HEK-293T cells stably expressing a dox-off Renilla were treated with 1 µg/mL doxycycline for 0–8 h. Reporter mRNA levels were measured at the indicated times using qPCR. Error bars are SD, n = 3. (B) Reporter mRNA stability analysis using Roadblock-qPCR. RNA was isolated from the same HEK-293T cells expressing Renilla mRNA from A, now treated with 4sU for 0–8 h, then treated with NEM and reverse transcribed. Reporter mRNA levels were measured using qPCR. Error bars are SD, n = 3.
