Figure 3 : Efficiency and fidelity of OEPR cloning for different sizes of homology arms.

From: OEPR Cloning: an Efficient and Seamless Cloning Strategy for Large- and Multi-Fragments

Figure 3

(A) Different homology arms (5, 10, 15, 20, 25, 30 and 35 bp (lanes 1–7, respectively)) were added to the 1 kb fragment from hNaV1.5. PCR product were detected by agarose gel electrophoresis. The arrow indicates the target band of the first PCR. (B) The purified fragments (containing 5, 10, 15, 20, 25, 30 and 35 bp homology arms (lanes 8–14, respectively)) from the 1st PCR were inserted into pGADT7 (7988 bp). PCR products were detected by agarose gel electrophoresis. The arrow indicates the target band of the second PCR. (C) Colony numbers per plate were counted to estimate cloning efficiencies. (D) The percentage of positive clones, which was estimated using colony PCR, was used to estimate cloning fidelities. Reported results are the mean ± SEM of three independent experiments. M: DNA molecular weight marker.