Figure 3From: Overcoming restriction as a barrier to DNA transformation in Caldicellulosiruptor species results in efficient marker replacementPCR analysis for the presence of pDCW89 after transformation into JWCB005. pDCW89 DNA isolated from C. hydrothermalis transformants was used to transform C. bescii (ΔpyrFA) and the presence of the plasmid was confirmed by PCR amplification of sequences contained only on the plasmid (A) the 0.9 kb PCR product of aac (Apramycin Resistance gene cassette), (B) the 1.3 kb PCR product of pyrF cassette and (C) the 1.6 kb PCR product of pSC101 ori region. Lane 1: total DNA isolated from JWCB005; lane 2: total DNA isolated from C. bescii transformant; lane 3: pDCW 89 isolated from E. coli. M: 1 kb DNA ladder (NEB).Back to article page