Antarctic Thermolabile UDG

Catalog #	Concentration	Size	List Price	Quantity	Your Price
M0372L	1000 units/ml	500 units	$510.00		$459.00
M0372S	1000 units/ml	100 units	$125.00		$112.50

Catalog #	Size	List Price	Your Price
M0372L	500 units	$510.00	$459.00
M0372S	100 units	$125.00	$112.50

Antarctic Thermolabile UDG (Uracil- DNA Glycosylase) catalyzes the release of free uracil from uracil-containing single-stranded or double-stranded DNA. The resulting abasic sites are susceptible to hydrolytic cleavage at elevated temperature and high pH. This enzyme is sensitive to heat and can be rapidly and completely inactivated at temperatures above 50°C.

To evaluate the capacity of carryover product digestion, a uracil-containing RT-qPCR product was generated using the Luna One-Step Probe RT-qPCR kit. Approximately 105 copies of the uracil-containing product were used as template for subsequent qPCR reactions using three different RT-qPCR master mixes (all containing UDG). Following manufacturer’s recommended protocols, incubation at 25˚C at the start of the 2nd reaction enables UDG to degrade the dU-containing input, reducing its ability to contribute to a (false) positive signal. The ΔC_q value is the cycle difference between carryover treatment and no carryover treatment of the same input. Larger ΔC_q values indicate more efficient carryover product digestion. After decontamination using the Luna mix, full product digestion was observed in one sample and a large ΔC_q was observed in the second. Note that the amount of DNA present in true contamination events is difficult to assess/predict.

Product Source

An E. coli strain that carries the cloned UDG gene from a psychrophilic marine bacterium

Reagents Supplied

The following reagents are supplied with this product:

NEB #

Component Name

Component #

Stored at (°C)

Amount

Concentration

M0372S -20

	Antarctic Thermolabile UDG	M0372SVIAL	-20	1 x 0.1 ml	1,000 units/ml
	Standard Taq Reaction Buffer Pack	B9014SVIAL	-20	1 x 1.5 ml	10 X

M0372L -20

	Antarctic Thermolabile UDG	M0372LVIAL	-20	1 x 0.5 ml	1,000 units/ml
	Standard Taq Reaction Buffer Pack	B9014SVIAL	-20	1 x 1.5 ml	10 X

Application Features

Prevention of Carry-over Contamination in PCR (1)
Remove Uracil-base from DNA

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 30°C in a total reaction volume of 50 µl in 1X Thermopol II Buffer.

Reaction Conditions

1X Standard Taq Reaction Buffer Pack
Incubate at 37°C

1X Standard Taq Reaction Buffer Pack
10 mM Tris-HCl
50 mM KCl
1.5 mM MgCl₂
(pH 8.3 @ 25°C)

Storage Buffer

10 mM Tris-HCl
50 mM NaCl
0.1 mM EDTA
1 mM DTT
50% Glycerol
pH 7.5 @ 25°C

Heat Inactivation

50°C for 5 minutes

Unit Assay Conditions

1X Taq 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.

Companion Products

Notes

One unit of enzyme is capable of converting 2.3 nmol of 5´ FAM-labeled 26-mer ssDNA with a single uracil to 10-mer ssDNA in 30 minutes at 37°C following NaOH and heat treatment. Activity is performed in a 50 μl standard Taq reaction buffer containing 2 pmol of 5´ FAM-labeled 26-mer ssDNA with a single uracil and variable amount of enzyme in 30 minutes at 37°C.
The NEB unit is 2–5 fold more active per unit than other suppliers. This Antarctic Thermolabile UDG is active in most PCR reaction buffers but is inhibited with increasing ionic strength (> 100 mM).

References

Longo, M.C. et al. (1990). Gene. 93, 125-128.

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FAQs

Citations

See more details on Bioz

Quality Control Assay

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.

Change Notifications

Effective 01/18/2024, unit definition 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 Data Sheets

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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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