Product Class: Other

Uracil-DNA Glycosylase (UDG)
NEBU cloned at NEB recombinant 37 No

UDG has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10225520. Learn more.

Product Introduction

Mechanism for M1282

  • Uracil DNA glycosylase
  • Monofunctional DNA glycosylase that catalyzes the hydrolysis of the N-glycosidic bond from deoxyuridine to release uracil
  • Active on ss and dsDNA
  • Heat inactivation not possible
Catalog # Size Concentration
M0280S 1000.0 units 5000 units/ml
M0280L 5000.0 units 5000 units/ml

Product Information

Description

E. coli Uracil-DNA Glycosylase (UDG) catalyses the release of free uracil from uracil-containing DNA. UDG efficiently hydrolyzes uracil from single-stranded or double-stranded DNA, but not from oligomers (6 or fewer bases).

Product Source

An E. coli strain that carries the UDG gene from E. coli.
This product is related to the following categories:

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • M0280S     -20    
      Uracil-DNA Glycosylase (UDG) M0280SVIAL -20 1 x 0.2 ml 5,000 units/ml
      UDG Reaction Buffer B0280SVIAL -20 1 x 1.5 ml 10 X
  • M0280L     -20    
      Uracil-DNA Glycosylase (UDG) M0280LVIAL -20 1 x 1 ml 5,000 units/ml
      UDG Reaction Buffer B0280SVIAL -20 1 x 1.5 ml 10 X

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to release 60 pmol per minute of a fluorescently labeled 47 mer single-stranded DNA oligonucleotide containing a single uracil base in 30 minutes at 37°C in a total reaction volume of 50 µl in 1X UDG Buffer.

Reaction Conditions

1X UDG Reaction Buffer
Incubate at 37°C

1X UDG Reaction Buffer
20 mM Tris-HCl
1 mM DTT
1 mM EDTA
(pH 8 @ 25°C)

Storage Buffer

10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
0.1 mg/ml Recombinant Albumin
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

No

Unit Assay Conditions

1X UDG Reaction Buffer, 1 unit of uracil DNA Glycosylase, 0.2 µg 3H-uracil DNA (104-105 cpm/μg) for 30 minutes at 37°C in a total reaction volume of 50 µl.

Application Features

  • Treatment of 0.1 µg of uracil-containing DNA with 1 unit of UDG for 10 minutes at 37°C renders the DNA incapable of being copied by DNA polymerase. The enzyme can be 95% heat killed by incubation at 95°C for 10 minutes. Since UDG remains partially active following heat treatment at 95°C, it is recommended that uracil glycosylase inhibitor be added to prevent degradation of product DNA. Alternatively, reaction products can be immediately extracted with phenol/chloroform.

Product Notes

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

References

  1. Lindahl, T. et al. (1977). J. Biol. Chem.. 252, 3286-3294.
  2. Wang, Z. et al. (1991). Gene. 99, 31-37.
  3. Devchand, P.R. et al. (1993). Nucl. Acids Res.. 21, 3437-3443.

Protocols, Manuals & Usage

Usage & Guidelines

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. What is the activity of UDG in the NEBuffers 1-4?
  2. Will UDG work in T4 DNA ligase buffer?
  3. What is the molecular weight of UDG?
  4. Is UDG a tagged protein?
  5. I see that UDG works optimally at 37°C. Do you have another glycosylase that works at higher temperatures?
  6. Does the Uracil Glycosylase inhibitor (UGI) inhibit UDG?
  7. Does UDG cut RNA?
  8. Can UDG be used to remove dU from a short 21mer oligo? Do the dU residues need to be spaced in any special way to avoid problems with cleavage?
  9. Are there any specific recommendations for the use of UDG on single stranded DNA? The materials on the web and data card seem to describe conditions for use with dsDNA.
  10. What is the difference between UDG and UNG?
  11. Does UDG release uracil from ss and dsDNA?

Quality, Safety & Legal

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specification Change Notifications

UDG has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10225520. All subsequent (higher number) lots will contain rAlbumin. For additional information, please visit at NEB Restriction Enzyme formulations with Recombinant Albumin (rAlbumin).

Effective 01/18/2024, unit definition has changed.

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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