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SSB was previously used to facilitate the analysis of target DNA using combination of PNA opener (ref. 3) and polymerase, in which SSB promoted strand-displacement by the polymerase;
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RecA protein catalyzes strand exchange between homologous single-stranded and double-stranded DNAs;
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The PNA strands bearing l-lysine residue at its C- and N-terminus were synthesized by Boc-chemistry, purified by the reversed-phase HPLC, and characterized by MALDI-TOF MS (Table S1 and Fig. S1, ESI). The target 203-mer dsDNA was prepared by PCR from pBR322 plasmid (C1724-G1926, Fig. S2, ESI). Gel-shift assay was achieved by 5%-nondenaturing polyacrylamide gel electrophoresis in TBE buffer at 25°C The weak band below the invasion complex in lane 4 (and lane 7) of Fig. 2 is associated with a non-specific complex between the dsDNA and SSB which is formed in equilibrium in the solutions. Consistently, this band is also observed in lane 3 where only dsDNA and SSB are mixed
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