Figure 5: OEPR cloning for assembly of multiple DNA fragments.
From: OEPR Cloning: an Efficient and Seamless Cloning Strategy for Large- and Multi-Fragments
(A) and (B) Multiple fragments of different sizes (two 1âkb, two 2âkb, two 3âkb, three 1âkb, three 2âkb and four 1âkb (lanes 1â6, respectively)) as well as 15âbp and 30âbp homology arms ((A) and (B), respectively) amplified from rNaV1.4 and hNaV1.5 were used as primers along with reverse primer R in 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. (C) Colony numbers per plates were determined to estimate cloning efficiencies for insertions of multiple fragments with 15âbp and 30âbp homology arms. (D) The percentages of positive clones, which were estimated by colony PCR, were used to estimate cloning fidelities. The results are the meanâÂąâSEM of three independent experiments. M: DNA molecular weight marker.
