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. Author manuscript; available in PMC: 2010 Apr 5.
Published in final edited form as: J Virol Methods. 2008 Sep 11;153(2):120–128. doi: 10.1016/j.jviromet.2008.07.024

Fig. 1.

Fig. 1

Generation of recombinant adenoviral plasmids in bacterial cells. (A) DNA from pShuttle-CMV-GFP was linearized by Pme I, dephosphorylated, and used for transformation of competent BJ5183 E. coli cells. Plasmid DNA prepared from inoculated colonies, as described in Section 2, were analyzed in supercoiled form by 0.4% agarose gel electrophoresis and ethidium bromide staining. Lane M, Fisher 1 kb DNA ladder (Cat. No. BP2553-100); lanes 1–38, plasmid DNA from different colonies. Based on the difference in migration rates, colonies in lanes 2, 3, 5, 6, 9–12, 14, 15, 17, 18, 22, 24–26, 28–30, 32, 33, 35, 37 contained self-ligated shuttle plasmid DNA; the remaining 15 out of 38 colonies potentially contained valid recombinant aden-oviral plasmids. (B) Analysis of plasmid DNA from post-recombination colonies. In addition to the treatments in (A), pShuttle-CMV-GFP DNA was treated with Taq DNA polymerase as described in Section 2. Based on the migration rates, none of the 38 colonies contained self-ligated shuttle plasmid DNA.