What is the optimal length of the homologous overlap between the termini of the PCR-amplified insert and linearized cloning vector?
What is the optimal length of the homologous overlap between the termini of the PCR-amplified insert and linearized cloning vector?
Current In?Fusion reaction conditions favor 15 bp of homologous overlap for single-insert cloning, and 20 bp of homologous overlap for multiple-insert cloning. We do not recommend using overlaps shorter than 12 bp or longer than 21 bp.