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PCR Master

plification of single-copy genes from eukaryotic genomes
  • Labeling of PCR products with modified nucleotides (DIG-dUTP, biotin-dUTP, fluorescein-dUTP)
  • Prevention of carry-over contamination, by incorporation of dUTP and degradation of contaminating DNA with Uracil-DNA Glycosylase
  • Cycle sequencing
  • Operating parameters
    • pH optimum: approx. 9 (adjusted at 20C)
    • Temperature optimum (for elongation): approx. 75C
      Note: The DNA polymerase has a half life at 95C of approx. 40 min
    • Divalent ion requirement: Mg2+ (standard concentration, 1.5 mM)
    • dNTP requirement: approx. 200 M for each dNTP
    Effects of additives Tween 20 (0.51.0%) detergent has been used to enhance the efficiency of Taq DNA Polymerase in certain PCRs. Other additives reported to have enhancing effects include DMSO, gelatine, glycerol, betaine, spermidine, T4 gene 32 protein, BSA, and ammonium sulfate. Key advantages
    • High PCR yield, because it is stable during prolonged, repetitive high temperature incubations
    • Enhances specificity of amplification, because it is highly processive and free of exonuclease and nicking activities
    • Increases sensitivity, because it produces good yields even when template amounts are limiting
    • Best results, because it has proved superior to competitors Taq DNA Polymerase preparations (see "Not all Taq DNA polymerase preparations are the same'"/>


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