VELOCITY PCR products are blunt-ended due to the 3'_5' exonuclease activity of the polymerase which removes 3'-A overhangs. A 3'-A overhang is useful, however, as it facilitates more efficient cloning into plasmid vectors and help prevent insert-to-insert ligation, eliminating possible tandem inserts. In order to generate a 3'-A overhang, the kit also contains a PCR Tailing Mix. This uses a uniquely blended Taq polymerase to add a single Adenine base as well as an exonuclease inhibitor to reduce the 3'_5' exonuclease activity of the VELOCITY, thus eliminating the need for purification of the PCR products prior to the addition of the overhang.
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