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PCR Master

for routine amplification of up to 3 kb genomic DNA targets Cat. No. 1 636 103 (100 reactions) Description The PCR Master Kit contains 10 vials of 2x concentrated PCR reaction mix (Taq DNA Polymerase, reaction buffer with MgCl2, stabilizing detergent, dNTPs) plus water PCR grade (for dilution), sufficient for 100 PCR reactions. Taq DNA Polymerase was originally isolated from the thermophilic eubacterium Thermus aquaticus BM. It is now supplied as a recombinant enzyme from E. coli. The enzyme is highly purified and is free of nonspecific endo- or exonucleases. Taq DNA Polymerase consists of a single polypeptide chain with a molecular weight of approximately 95 kD. It is a highly processive 5'3' DNA polymerase which lacks 3'5' exonuclease activity.

Note: For additional convenience and reliable PCR with minimum reagent preparation, we offer the PCR Master [Cat. No. 1636103, 2x conc. PCR reaction mix (Taq DNA Polymerase, reaction buffer with MgCl2, stabilising detergent, dNTPs)] plus water PCR grade for dilution. Further the PCR Core Kit (Cat. No. 1578553), containing Taq DNA Polymerase, dNTP stock solution, PCR reaction buffer with and without MgCl2 and a MgCl2 stock solution. Application Amplifies genomic targets up to 3 kb with good yield and specificity. Capable of amplifying lambda DNA up to 10 kb. The enzyme is ideal for:
  • Routine am 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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