5' RACE Protocol for Bacterial Genes

5’ RACE was performed using a 5’/3’ RACE kit (Roche, Germany) as previously described [62 (link)]. Essentially, DNase I-treated total RNA was reverse transcribed using specific primers BT3311 and BT3337 as SP1 primers for finR and fprA, respectively. The first-strand DNA (cDNA) was purified, and poly(A) was added to the 5’-terminus of the cDNA using terminal transferase. Next, poly(A)-tailed cDNA was PCR-amplified using the specific SP2 primer BT4438 for finR and BT4479 for fprA and an anchored oligo(dT) primer. The purified PCR product was cloned into the pGemT vector, and the +1 site was identified from the DNA sequences.