Fig. 1

Schematic representation of MRF cloning. The insert gene or vector was amplified by regular double-primer PCR using genomic DNA, cDNA, or the original vector as template. Compatible cohesive ends of insert gene or vector were created by two single-primer linear PCRs performed in parallel, followed by annealing of the two PCR products. Inserts and acceptors with compatible cohesive ends were then assembled by ligation