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Applied Biosystems™ AmpErase™ Uracil N-Glycosylase (UNG)

Catalog No. N8080096
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N8080096 100 units
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Catalog No. N8080096 Supplier Applied Biosystems™ Supplier No. N8080096
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Description

Includes

6 x 1.5mL vial each of GeneAmp 10X PCR Buffer II and MgCl2 sufficient for 100 PCR amplifications of 100μL each.

Enzyme removes any uracil incorporated into single- or double-stranded DNA

AmpErase™ Uracil N-Glycosylase (UNG), part of GeneAmp™ PCR Carry-over Prevention Kit, is 26 kDa ultrapure, recombinant enzyme encoded by E. coli uracil N-glycosylase gene, inserted into E. coli host to direct expression of wild type form of enzyme. Enzyme removes any uracil incorporated into single- or double-stranded DNA.

  • Enzymatic method stops PCR carryover contamination, which prevents false positive result
  • No interference with any PCR or real-time PCR application
  • Optimized for use with GeneAmp™ PCR core reagents and GeneAmp™ instrument systems

PCR, PCR and Real-Time PCR, Real Time PCR (qPCR)

Order Info

Shipping Condition: Dry ice

Specifications

Content And Storage Contains AmpErase™ Uracil N-glycosylase (UNG) (100 units). Sufficient for 100 PCR amplifications, 100 μl each. Store at -20°C.
PCR Method qPCR, Standard PCR
Polymerase DNA Polymerase
Product Line AmpErase
Product Type Uracil N-Glycosylase for PCR reactions
Quantity 100 units
Shipping Condition Dry Ice
Sufficient For 100 PCR Amplifications
For Use With (Application) Standard PCR

Frequently Asked Questions (FAQs)

What is AmpErase UNG, and how does it work?

AmpErase UNG (Uracil N-glycosylase) is an enzyme utilized in a powerful method for elimination of carryover PCR products in Real-Time PCR. This method modifies PCR products such that in a new reaction, any residual products from previous PCR amplifications will be digested and prevented from amplifying, but the true DNA templates will be unaffected.

Here is how it works: During amplification, dUTP is substituted for dTTP, resulting in dUTP-containing amplicons. In subsequent reactions, a short pre-PCR incubation step will allow the AmpErase UNG to digest any dUTP containing DNA. Since AmpErase UNG is active on both single- and double-stranded dUTP-containing DNA, the procedure should work on dU-containing PCR products from standard or asymmetric PCR amplifications. However, uracil ribonucleotide residues in RNA, novel DNA containing dTTP, or cDNA containing dTTP will not be suitable substrates for UNG, so your templates will be unaffected.

Note that this is a proactive method to prevent contamination from future reactions, but will not help with a pre-existing contamination problem of standard dTTP-containing PCR products. That can only be remedied with thorough cleaning of lab surfaces, equipment and air filters.

I am using AmpErase Uracil N-glycosylase (UNG) and dUTP in my PCR reactions and would like to use the PCR product in a post-PCR application. Does the dUTP affect my ability to hybridize, sequence, clone, or digest the PCR product?

PCR products containing dUTP residues are generally equivalent to dT-containing PCR products as hybridization targets and as templates for dideoxy-terminated sequencing reactions. They can also work equivalently as targets for direct cloning as long as they are transferred into UNG-minus bacterial hosts.

The recognition of dU-containing DNA by restriction endonucleases has been studied, and depending on the specific endonuclease there may be no effect on enzymatic activity on the substitution of dU for dT (e.g., EcoR1 and BamH1), or the dU-containing DNA is cleaved more slowly than dT-containing DNA (e.g., Hpa1, HindII, and HindIII). The most accurate answer on the effects of substituting dU for dT are best determined using your own empirical studies.

For Research Use Only. Not for use in diagnostic procedures.