Fig. 5From: AFEAP cloning: a precise and efficient method for large DNA sequence assemblyConstruction of larger plasmid. a Schematic diagram of the assembly of 200 kb BAC with AFEAP cloning method. (b, upper panel) Agarose electrophoresis shows the PCR amplification using the primers as listed at Additional file 2: Table S2. Lane 1: 1 kb DNA ladder; Lane 2–9: PCR products from first-round PCRs; Lane 10: Annealing of PCR products from second-round PCRs before ligation; Lane 11: after ligation. DNA samples were electrophoresed in 1% agarose gel. (b, below panel) Sequencing validation of the re-joining junction sites. The overhang regions are marked by red boxesBack to article page