Figure 1. Three-step PCR reactions to assemble linear expression template.

(A) Schematic representation of PCR-based generation of fusion constructs. Two primary PCR products with defined overlapping ends are synthesized by the first PCR reaction. These two fragments are joined in a second PCR, overlap extension PCR and subsequent third PCR step introduces the regulatory elements necessary for transcription and translation to the fused target genes. (B) Sequence elements of final amplified expression template.