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Glycerol-Free Heat-labile UDGase (HC)

Glycerol-Free Heat-labile Uracil DNA Glycosylase (UDG) is a high concentration enzyme used to eliminate carryover polymerase chain reaction (PCR) products in qPCR and RT-qPCR. This is done by incorporating dUTP in all qPCR products, so any residual products from previous PCR amplifications will contain dUs and can be digested by pre-treating with UDG enzyme.


*Not available for sale in the United States

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Meridian's Glycerol-Free Heat-labile UDGase (HC)

  • Optimized dTTP to dUTP ratio for high sensitivity and specificity.

  • High efficiency, giving excellent performance in multiplex qPCR assays.

  • Prevents carry-over qPCR contamination.

  • UDGase is also available as glycerol-free, high concentration.

High efficiency and sensitivity with DNA viruses

Uracil DNA Glycosylase graph
Amplification of a 10-fold serial dilution of Parvovirus B19 from inactivated crude viral lysates was tested using Hi-throughput dUTP qPCR Mix (MDX031) and primers and probe for Parvovirus B19 and Uracil DNA Glycosylase (MDX054). Four replicated are shown in the amplification curve, illustrate the reproducibility, efficiency and sensitivity of Hi-throughput dUTP qPCR Mix.

Glycerol-Free Heat-labile UDGase (HC), MDX217

Available in 20 µL (1000 Units) and 500µL (25,000 Units) aliquots

Description

Uracil DNA Glycosylase (UDG) catalyzes the release of uracil from uracil-containing single or double-stranded DNA, but not from RNA or oligonucleotides (6 or fewer bases). UDG is active over a broad pH range with an optimum at pH 8.0, does not require a divalent cation, and is inhibited by high ionic strength (>200 mM). Glycerol-Free Heat-labile UDGase (HC), derived from Gadus morhua has all the attributes of the enzyme derived from E. coli with the added benefit of being heat-labile. It is completely and irreversibly inactivated after 10-minute incubation at 50°C.




Specifications

Description Uracil DNA Glycosylase (UDG) removes uracil residues from the sugar moiety of single- and double-stranded DNA without destroying the phosphodiester backbone, preventing its use as a hybridization target or as a template for DNA polymerases. UDG will not remove uracil from RNA.
Concentration 20 U/µL
Unit Definition One unit is the amount of enzyme that catalyses the release of 60 pmol of uracil per minute from double stranded, uracil-containing DNA. Activity was measured by release of [3H]-uracil in a 50 µL reaction mix containing 0.2 mg DNA (104-105 cpm/mg) in 30 minutes at 37 °C.
Source MDX217 derived from Gadus morhua, has all the attributes of the enzyme derived from E. coli with the added benefit of being heat labile.
Format Clear, colorless solution
10x reaction buffer 0.25 M Tris-Cl, 1 mM EDTA, 10 mM DTT, pH 8.0.
Application High throughput, PCR, qPCR
Sample type cDNA, DNA
Presentation 2 vials
Storage -20 °C
Mix stability See outer label
Functionality dUTP PCR products are degraded on the addition of UDGase to give single, distinct bands on an agarose gel.
DNA Contamination None detected in PCR amplification with traces overlay with the negative control on E. coli and mouse genomic DNA specific targets.
DNase Contamination No detectable degradation

FAQs: Glycerol-Free Heat-labile UDGase (HC)

During PCR, misincorporation of uracil during DNA synthesis or spontaneous cytosine deamination in the DNA strand can occur. UDG enzyme catalyzes the excision of uracil nucleobases (dUTPs) to prevent mutagenesis, allowing for targeted, more reliable PCR assays.

UDG refers to a superfamily of enzymes that includes family I UDG enzyme called UNG, after the uracil-N-glycosylase gene, so the terms UDG enzyme (UDGase) and UNG enzyme are commonly used interchangeably because they perform the same function in qPCR, by preventing carryover contamination.

Uracil DNA Glycosylase (UDG) catalyzes the release of uracil from uracil-containing single or double-stranded DNA, but not from RNA or oligonucleotides (6 or fewer bases).

UDG is active over a broad pH range with an optimum at pH 8.0, does not require a divalent cation, and is inhibited by high ionic strength (>200 mM).

The Uracil DNA Glycosylase (MDX054) and Glycerol-Free UDGase (HC) (MDX216) enzyme has activity below 55°C, so this should be the minimum annealing temperature for qPCR amplification, to avoid degradation of newly synthesized dU-containing qPCR products, the enzyme isirreversibly inactivated by incubation at 95°C for 10 min. For Glycerol-Free Heat-labile UDGase (HC) (MDX217), the enzyme is irreversibly inactivated by incubation at 50°C for 10 min.

Uracil DNA Glycosylase is tested for purity (run on SDS-PAGE gel), presence and absence of endonucleases, nickases, exonucleases and RNases.

The Uracil DNA Glycosylase (MDX054) and Glycerol-Free UDGase (HC) (MDX216) is sourced from a recombinant E. coli strain carrying the over-expressed modified UDG gene. The Glycerol-Free Heat-labile UDGase (HC) (MDX217) is sourced from Gadus morhua.

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