Product Class: Other

Thermostable Endonuclease Q
neb31 cloned at NEB recombinant 65 No


Catalog #M0701

Product Introduction

E1604 Time Temp

  • DNA endonuclease
  • Active on ssDNA and dsDNA
  • Catalyzes the cleavage of a phosphodiester bond 5′ from deoxyxanthosine (dX), deoxyinosine (dI), deoxyuracil (dU), and an abasic (AP) site, creating a nick with 3′-OH and a 5′-phosphate
  • Thermostable

Product Information

Description

Thermostable Endonuclease Q (EndoQ) is an archaeal DNA endonuclease that cleaves the phosphate backbone of a DNA substrate 5′ to the position of the modified nucleobases of deoxyxanthosine (dX), deoxyinosine (dI), deoxyuracil (dU), and an abasic (AP) site. The result is a 3′-OH and a 5′-phosphate. Thermostable Endonuclease Q generally prefers single-stranded DNA over double-stranded DNA, and exhibits higher activity on dU over dI and dX.

Product Source: An E. coli strain that carries a cloned archaeal Endonuclease Q gene with a C-terminal his-tag.


Figure 1: Relative Activity of Thermostable Endonuclease Q on Various DNA Substrates

Relative Activity of Thermostable Endonuclease Q

Thermostable Endonuclease Q has different specificities for modified nucleotides in ssDNA and dsDNA. Thermostable Endonuclease Q (#M0701) activity was tested on 1 pmol of ssDNA or dsDNA substrates with fluorescently-labeled oligonucleotides containing modified nucleotides of interest (red) for 1 hour in 1x NEBuffer r2.1 at 65°C. Cleaved products were detected by capillary electrophoresis. Thermostable Endonuclease Q prefers ssDNA, and exhibits higher activity on dU over dI and dX.

Figure 2: Temperature Profile of Thermostable Endonuclease Q

Temperature Profile of Thermostable Endonuclease Q
Thermostable Endonuclease Q is active over a wide range of temperatures. Thermostable Endonuclease Q (#M0701, 0.2 U) activity was tested on 1 pmol of a fluorescently-labeled single-stranded oligonucleotide containing a single uracil for 1 hour in 1x NEBuffer r2.1 at various temperatures. Cleaved products were detected by capillary electrophoresis. Thermostable Endonuclease Q is active over a wide range of temperatures (37°C – 90°C), but displays optimal activity between 65°C and 85°C.

Figure 3: Thermostable Endonuclease Q is active in some DNA:RNA contexts

Thermostable Endonuclease Q active in some DNA:RNA
Thermostable Endonuclease Q is active on some DNA:RNA contexts. Thermostable Endonuclease Q (#M0701, 0.2 U) activity was tested on 1 pmol of substrate containing a single deoxyuracil (dU) or ribouracil (rU) within a DNA oligonucleotide (gray) or RNA oligonucleotide (blue) for 1 hour in 1x ThermoPol® Reaction Buffer. Activity was compared with Thermostable USER® III Enzyme (#M5509) for comparison. Cleaved products were detected by capillary electrophoresis. Thermostable Endonuclease Q can cleave at dU and rU located within DNA oligonucleotide strands regardless of presence or type (DNA or RNA) of complement.
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

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to cleave 1 pmol of a 60-mer oligonucleotide ssDNA containing a single deoxyxanthosine site* in a total reaction volume of 20 µl after 1 hour at 65°C.  

*A deoxyxanthosine site is synthetically prepared with dX at the 24th position of a 5´ FAM-labelled 60-mer ssDNA oligo.

Reaction Conditions

1X NEBuffer™ r2.1
Incubate at 65°C

1X NEBuffer™ r2.1
50 mM NaCl
10 mM Tris-HCl
10 mM MgCl2
100 µg/ml Recombinant Albumin
(pH 7.9 @ 25°C)

Storage Buffer

10 mM Tris-HCl
500 mM NaCl
0.1 mM EDTA
1 mM DTT
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

No

Protocols, Manuals & Usage

Protocols

  1. Protocol for Thermostable Endonuclease Q (NEB #M0701)

Usage & Guidelines

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. What is the difference between the suite of USER® enzymes (#M5505, #M5508, #M5509) and Thermostable Endonuclease Q (#M0701)?
  2. What is the difference between Endonuclease V (#M0305) and Thermostable Endonuclease Q (#M0701)?
  3. What is the activity of Thermostable Endonuclease Q (#M0701) in different NEBuffers?
  4. What is the molecular weight of Thermostable Endonuclease Q (#M0701)?
  5. Is Thermostable Endonuclease Q (#M0701) a tagged protein?
  6. Can the product of the Thermostable Endonuclease Q (#M0701) reaction be ligated?
  7. How close to the 5′ DNA end will Thermostable Endonuclease Q (#M0701) cleave?
  8. How close to the 3’ DNA end will Thermostable Endonuclease Q (#M0701) cleave?
  9. What temperatures is Thermostable Endonuclease Q (#M0701) active?
  10. What modified bases will Thermostable Endonuclease Q (#M0701) recognize and cleave?
  11. Does Thermostable Endonuclease Q (#M0701) have activity in DNA-RNA hybrid duplexes?
  12. What is the relative activity of Thermostable Endonuclease Q (#M0701) on different modified nucleobase-containing substrates?

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.

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.

Other Products You May Be Interested In