GoldStar® DNA polymerase is purified from an E. coli strain, which carries a Thermus species (new strain) DNA polymerase overproducing plasmid. Activity
The GoldStar® show svery good fidelity and an average error rate around 5.10
-5.
The GoldStar® DNA polymerase has 5’ > 3’ exonuclease activity but lacks 3’ > 5’ proofreading activity. It also has the extendase activity allowing TA cloning.
Application
DNA fragments as long as 12 kb can be efficiently amplified in the presence of inhibiting impurities still present in the DNA (i.e. cell lysate material). The amplification is efficient and the results reproducible using standard PCR conditions and total genomic DNA. Improvement can be obtained by increasing the magnesium concentration and extension time, and by the use of longer primers.
A concentration higher than 1.5 mM Mg
2+ is required for DNA fragments > 5 kb. Since Mg
2+ stabilizes the DNA double strand, excessive amounts will prevent a complete denaturation. This reduces the extension yield and leads to spurious primer / template annealing, thus decreasing specificity.