Figure 5.

PCR amplification of ~5 kb of DNA in pET17b[Pfu-Pol] by polymerase variants. Reactions were carried out using the polymerase variants indicated (a = 20 nM; b = 100 nM) in two buffer systems (A = Tris-HCl pH 8; B = Bicine-NaOH pH 9). Analysis was by 1% agarose gel electrophoresis with ethidium bromide staining. The standards comprise a 1 kb ladder with the more intense bands at 1, 3, and 6 kB. The expected PCR product (~5 kb) is indicated with the arrow.