Exonuclease T

Catalog #	Concentration	Size	List Price	Quantity	Your Price
M0265L	5000 units/ml	1250 units	$483.00		$434.70
M0265S	5000 units/ml	250 units	$119.00		$107.10

Catalog #	Size	List Price	Your Price
M0265L	1250 units	$483.00	$434.70
M0265S	250 units	$119.00	$107.10

Exonuclease T (Exo T), also known as RNase T, is a single-stranded RNA (1,2) or DNA (3,4) specific nuclease that requires a free 3´ terminus and removes nucleotides in the 3´→ 5´ direction. Exonuclease T can be used to generate blunt ends from RNA (5) or DNA molecules that have 3´ extensions (4).

Product Source

Exonuclease T is overexpressed and purified as a C-terminal fusion to maltose-binding protein (MBP). MBP is removed from Exonuclease T by Factor Xa cleavage and Exonuclease T is then purified away from Factor Xa and MBP. Exonuclease T cleaved from MBP has an additional amino acid on the N-terminus and a Phe instead of a Met (i.e., Glu-Phe-Exo T instead of Met-Exo T).

Reagents Supplied

The following reagents are supplied with this product:

NEB #

Component Name

Component #

Stored at (°C)

Amount

Concentration

M0265S -20

	Exonuclease T	M0265SVIAL	-20	1 x 0.05 ml	5,000 units/ml
	NEBuffer™ 4	B7004SVIAL	-20	1 x 1.25 ml	10 X

M0265L -20

	Exonuclease T	M0265LVIAL	-20	1 x 0.25 ml	5,000 units/ml
	NEBuffer™ 4	B7004SVIAL	-20	1 x 1.25 ml	10 X

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to release 1 nmol of single dT nucleotides from 50 pmol of Fam-labeled polythymidine substrate in 30 minutes at 25°C.

Reaction Conditions

1X NEBuffer™ 4
Incubate at 25°C

1X NEBuffer™ 4
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
1 mM DTT
(pH 7.9 @ 25°C)

Activity in NEBuffers

NEBuffer™ 1: 100%
NEBuffer™ 2: 75%
NEBuffer™ 3: 25%

Storage Buffer

10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
200 Âµg/ml BSA
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

65°C for 20 minutes

Materials Sold Separately

NEBuffer 4

Notes

Exonuclease T has a different activity on RNA vs. DNA. For RNA, 1 unit of Exonuclease T is required to completely digest 1.0 pmol of rA20 under standard reaction conditions as measured by gel electrophoresis.
Single stranded RNA molecules containing C residues at its 3'-terminus might be difficult to digest with Exonuclease T.
Exonuclease T removes ssDNA extensions but does not work well when two cytosine nucleotides are in a row. “Although RNase T appears to bind up to 10 nucleotides in its active site, its specificity is defined largely by the last 4 residues. A single 3'-terminal C residue can reduce RNase T action by >100-fold, and 2-terminal C residues essentially stop the enzyme.” (PMID: 12050169 DOI: 10.1074/jbc.M204252200)

References

Deutscher, M. P., Marlor, C. W. and Zaniewski, R. (1984). Proc. Natl. Acad. Sci. USA. 81, 4290–4293.
Deutscher, M. P. and Marlor, C. W. (1985). J. Biol. Chem. 260, 7067–7071.
Viswanathan, M., Dower, K. D. and Lovett, S. T. (1998). J. Biol. Chem. 273, 35126–35131.
Zuo, Y. and Deutscher, M. P. (1999). Nucleic Acid Res. 27, 4077–4082.
Zeng, Y. and Cullen, B. R. (2004). Nucleic Acid Res. 32, 4776–4780.
Zuo, Y. and Deutscher, M.P. (2002). J. Biol. Chem. 277, 29654-61.

Selection Charts

Web Tools

Exo Selector

Citations

See more details on Bioz

Additional Citations

Zuo Y, Deutscher MP. (2002) The physiological role of RNase T can be explained by its unusual substrate specificity. J Biol Chem 277(33):29654‐29661. DOI: 10.1074/jbc.M204252200

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.

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]

M0265S_L_v1

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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