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. 2013 Apr 5;41(10):5303–5320. doi: 10.1093/nar/gkt207

Figure 7.

Figure 7.

DnaEBS extends RNA primers after de novo di-ribonucleotide polymerization by DnaG. (A) Representative chromatograms from primase assays showing RNA primers synthesized by B. subtilis DnaG (1.8 µM) in reactions containing the 23-mer 5′-d(CTA)-specific ssDNA template and 0.8 mM each NTP before RNase H treatment. (B) RNA primers synthesized as described in panel A were incubated with RNase H to confirm content. (C) Priming reactions included dNTPs without the presence of ribonucleotides. (D) Priming reactions contained DnaEBs, as well as both NTPs and dNTPs. (E) B. subtilis DnaC helicase was added to the priming reactions, along with DnaEBs and both NTPs and dNTPs. (F) RNA:DNA hybrid primers synthesized in the reaction described in panel D were treated with RNase H to determine primer composition. Elution times for the various primer products synthesized were determined by using RNA, DNA and RNA:DNA size markers shown in Supplementary Table S2. A schematic of de novo primer synthesis and extension is shown below the chromatograms.