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. 2010 Nov 24;5(11):e14112. doi: 10.1371/journal.pone.0014112

Figure 4. RT-PCR detection of sodC mRNA in B. neotomae and B. abortus RB51 (A) and identification of their 5′ ends by RACE analysis (B).

Figure 4

A) Total RNA extracted from the bacterial cultures at late log phase was used as template in RT-PCR or direct PCR for amplification of a 5′ portion of sodC mRNA using a pair of specific primers. Post-amplification reaction mixtures of tubes containing RNA extracted from B. neotomae (lanes 1 and 2) or B. abortus RB51 (lanes 3 and 4) were separated on a 2% agarose gel by electrophoresis and stained with ethidium bromide to visualize the DNA fragments. Lanes 1 and 3, RT-PCR reaction mixtures. Lanes 2 and 4, PCR reaction mixtures. Lanes 5 and 6, reaction mixtures from no-template-controls of RT-PCR and PCR reactions, respectively. Lane marked MW contains DNA molecular size marker, and the numbers at the left indicate molecular size in base-pairs. B) Representative nucleotide sequencing chromatograms showing the location of 5′ ends of the cloned sodC cDNAs from B. abortus RB51 (i & ii) and B. neotomae (ii).