Figure 5. Pfu was used to efficiently amplify repeated sequences by PCR.

(A) Taq-based PCR was unsuitable for screening multiple insertions as it produced strong intermediate-sized products and weak specific-products when amplifying templates containing 1 to 7 hairpin expression cassettes. (B) A series of approximately equivalent sized plasmids with non-structured (& non-repeated) inserts was successfully amplified with Taq. (C) Taq was unsuitable for amplifying templates containing 1 to 7 promoter-only cassettes (no hairpin sequences). (D) Several different polymerases (Phusion, Dynazyme EXT, Dynazyme II, Immolase and Pfu) were tested with the promoter-only series of vectors using the manufacturers recommended starting conditions.