Total RNA from exponentially growing plasmid-carrying E. coli strains was extracted (41 (link)). Total RNA (10 to 20 µg) was hybridized with 5′-32P-labeled ML-512 and ML-1314 primers. Reverse transcription experiments were done at 42°C using the reverse transcriptase ImProm-II (Promega). Reaction products were analyzed on an 8% polyacrylamide gel in parallel with sequencing reaction products obtained using the same primers.
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