DAp GoldStar® is a high-performance proprietary complex of enzymes and additives, specially designed for applications requiring high processivity and high yields. It is recommended for difficult, long, and / or impure fragments. An advanced incubation buffer is supplied for optimal results and enhance enzyme life. The HiSpec buffer improves specificity of a reaction, especially on GC-rich or difficult templates.
Activity
DAp GoldStar® has 5’ ’ 3’ exonuclease and posseses 3’ ’ 5’ proofreading activity, which prevents misincorporations during extention. It also has the ”extendase“ activity allowing TA cloning.
Application
Long Region Applications: Optimal composition of different enzymatic activities enables DAp GoldStar® to perform primer extension over long regions by reducing premature strand termination and template degradation. Using long primers at elevated Mg2+ concentrations, > 30 kb or 15 kb products can be achieved from lambda templates or genomic DNA respectively.
High Performance: DAp GoldStar® provides high yield and specificity, even with impure DNA or difficult templates. Unlike standard 3’ ’ 5’ proof-reading polymerases, DAp GoldStar® can be used in combination with primers containing wobbles.
Direct Cloning: DAp GoldStar® is recommended for direct gene cloning without the need to verify the sequence prior to expression. DAp GoldStar® exhibits the necessary terminal transferase activity for effective integration of primer extension products into TA cloning vectors, even from difficult templates.
High Fidelity: DAp GoldStar® is a mix of polymerases that possesses 5’ ’ 3’ DNA polymerase activity and 3’ ’ 5’ proofreading activity which prevents misincorporations during primer extension. DAp GoldStar® contains an additive, which eliminates the sequence-dependent removal of 3’-terminal dNMPs in growing DNA chains (pyrophosphoro-lysis). This combination of properties can provide a > 7 fold higher fidelity than Taq DNA polymerase. In contrast with other proofreading enzymes, DAp GoldStar® does not degrade primers.
Note: For long regions, the addition of glycerol or DMSO to a final concentration of 2-8 % may improve performance.