Figure 4: Large fragment insertion by OEPR cloning.
From: OEPR Cloning: an Efficient and Seamless Cloning Strategy for Large- and Multi-Fragments
(A) and (B) Large fragments of various lengths (1, 2, 3, 4, 5 and 6âkb (lanes 1â6, respectively)) containing 15âbp and 30âbp homology arms ((A) and (B), respectively) amplified from hNaV1.5 were used along with reverse primer R as primers for the second PCR step. The second PCR product (5âÎźl) was detected by agarose gel electrophoresis. The white arrow indicates the target band of the final product after the second of PCR. Different dosages of insert fragments using 15âbp (C) and 30âbp (D) homology arms in the first PCR were step were examined. Colony numbers on plates were counted to estimate cloning efficiencies. (E) Percentages of positive clones, which were estimated by colony PCR, were determined to estimate cloning fidelities. Reported results are the meanâÂąâSEM of three independent experiments. M: DNA molecular weight marker.
