Product Catalog

U-DNA cleavage can be used to increase the efficiency of site-directed mutagenesis. For this purpose, uracil is incorporated in vivo into the target DNA. After primer annealing, and enzymatic filling-in, and ligation, the selection for the mutated strand is performed by enzymatic degradation of the original non-mutated uracil, which contains the wild-type strand, using UNG. A further application is carryover prevention for PCR. During PCR, dUTP can be incorporated into the PCR product. Before a subsequent amplification reaction, contaminating PCR products are degraded using uracil-DNA glycosylase. The enzyme is inactivated at the beginning of the amplification reaction by heating to 95°C. Simultaneous strand cleavage of the U-DNA is achieved during this heating step.

The enzyme is supplied in storage buffer, 1 U/µl.

Function test: Carryover prevention activity is assayed by adding approximately 10⁵ dU that contains templates prior to the amplification reaction. After UNG treatment, no amplification products could be detected. The enzyme does not contain any contaminating exo- or endonucleases.

Associated Products

• Taq DNA Polymerase, 5 U/μl;
  from Thermus aquaticus BM, recombinant (E. coli) Thermostable DNA Polymerase
• PCR Nucleotide Mix^PLUS;
  Premixed deoxynucleotide solution for use in PCR and RT-PCR carryover prevention
• Taq DNA Polymerase, dNTPack;
  5 U/μl Taq DNA Polymerase with ready-to-use PCR-Grade Nucleotide Mix
• Expand High Fidelity^PLUS PCR System;
  Thermostable DNA polymerase blend
• Expand High Fidelity^PLUS PCR System, dNTPack;
• Uracil-DNA Glycosylase, heat-labile;
  from marine bacterium BMTU 3346
• PCR Core Kit^PLUS;
  Kit for PCR and PCR carryover prevention
• dUTP;
  PCR Grade, Na-Salt
• Agarose Gel DNA Extraction Kit;
  For the elution of DNA fragments from agarose gels
• High Pure PCR Product Purification Kit;
  For purification of PCR reaction products
• High Pure PCR Template Preparation Kit;
  For isolation of genomic nucleic acids for PCR, restriction-enzyme analysis, and Southern blotting