RedGoldStar DNA polymerase is purified from an E. coli strain, which carries a Thermus species (new strain) DNA polymerase overproducing plasmid.
The RedGoldstar is the GoldStar® DNA polymerase version containing a red dye. The dye does not influence the PCR reaction and does not fluoresce. RedGoldStar DNA polymerase allows to visually check the presence of polymerase in the reaction mixture.
RedGoldStar is a recombinant Taq DNA polymerases that is isolated from extremely thermostable clones.
Activity
The RedGoldStar show svery good fidelity and an average error rate around 5.10-5.
The RedGoldStar DNA polymerases has 5’ > 3’ exonuclease activity but lack 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 Mg2+ is required for DNA fragments > 5 kb. Since Mg2+ 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.