Product Class: Restriction Endonuclease

Nt.BstNBI
neb31 cloned at NEB recombinant dil_A 55 80 Heat

Nt.BstNBI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10159526. Learn more.

Now available in WarmStart version (NEB #R0725) which allows for RT set up of SDA.

This is product is available in a glycerol-free format. Contact us for more information.

We are excited to announce that all reaction buffers are now BSA-free. NEB began switching our BSA-containing reaction buffers in April 2021 to buffers containing Recombinant Albumin (rAlbumin) for restriction enzymes and some DNA modifying enzymes. Find more details at www.neb.com/BSA-free.


NEB restriction endonuclease that recognizes the sequence GAGTCNNNN^N
Isoschizomers | Single Letter Code

Product Introduction

  • This is a nicking endonuclease
  • Generates DNA molecules that are “nicked”, rather than cleaved
  • Nicking Enzyme Cut Site: GAGTC (4/none)
Catalog # Size Concentration
R0607S 1000.0 units 10000 units/ml
R0607L 5000.0 units 10000 units/ml

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Product Information

Description

Highlights

  • N.BstNB I catalyzes a single-stranded nick 3' of its recognition sequence.
Nt.BstNBI is a site specific endonuclease that cleaves only one strand of DNA on a double-stranded DNA substrate. The nicking endonuclease catalyzes a single strand break 4 bases beyond the 3´ side of the recognition sequence.

Product Source

An E. coli strain that carries the cloned Nt.BstNBI gene from Bacillus stereothermophilus 33M (Z. Chen).
This product is related to the following categories:
Nicking Endonucleases Products,
Restriction Endonucleases: N-O Products,
This product can be used in the following applications:
Strand Displacement Amplification & Nicking Enzyme

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 digest 1 µg T7 DNA in 1 hour at 55°C in a total reaction volume of 50 µl.

The phage T7 DNA is a particularly convenient substrate for assaying the activity because of the proximity of GAGTC sequences. Nt.BstNBI cleavage of phage T7 DNA yields 5 bands; 13 kb, 9 kb, 7 kb, 6 kb and 4 kb.

Reaction Conditions

1X NEBuffer™ r3.1
Incubate at 55°C

1X NEBuffer™ r3.1
100 mM NaCl
50 mM Tris-HCl
10 mM MgCl2
100 µg/ml Recombinant Albumin
(pH 7.9 @ 25°C)

Activity in NEBuffers

NEBuffer™ r1.1: 0%
NEBuffer™ r2.1: 10%
NEBuffer™ r3.1: 100%
rCutSmart™ Buffer: 10%

Diluent Compatibility

Storage Buffer

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

Heat Inactivation

80°C for 20 minutes

Methylation Sensitivity

dam methylation: Not Sensitive
dcm methylation: Not Sensitive
CpG Methylation: Not Sensitive

Activity at Temperature

@37°C: 50%

Product Notes

  1. To see how this enzyme (along with other nicking enzymes) performs at different temperatures, please visit Effect of Various Temperatures of Different Nicking Endonucleases.
  2. Not sensitive to CpG, dcm, or dam methylation.

References

  1. Song, Q. et al. (2010). Anal.Chem. [Epub ahead of print].
  2. Zhang, P. et al. (2010). ProteinExpr. Purif. 69, 226-234. [Epub 2009 Sep 9].

Protocols, Manuals & Usage

Protocols

  1. Optimizing Restriction Endonuclease Reactions
  2. Restriction Digest Protocol
  3. Double Digest Protocol with Standard Restriction Enzymes

Usage & Guidelines

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. What is the activity of nicking enzymes at different temperatures?
  2. Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
  3. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?

Troubleshooting

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

Nt.BstNBI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10159526. All subsequent (higher number) lots will contain rAlbumin. For additional information, please visit at NEB Restriction Enzyme formulations with Recombinant Albumin (rAlbumin).

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 [email protected].

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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