Product Class: Restriction Endonuclease

Nt.CviPII
neb31 cloned at NEB recombinant dil_A 37 65 Heat CpG

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 ^CCD

Product Introduction

  • Nt.CviPII is used in the NicE-Seq protocol, which is used for high resolution open chromatin profiling. Please refer to FAQs and protocols (below) for more information.
  • This is a nicking endonuclease, which generates DNA molecules that are “nicked”, rather than cleaved
  • Nicking Enzyme Cut Site: CCD (-3/none)
Catalog # Size Concentration
R0626S 40.0 units 2000 units/ml

Product Information

Description

Nt.CviPII is a nicking endonuclease that cleaves only one strand of DNA on a double-stranded DNA substrate. The final product on pUC19 is an array of bands from 25 to 200 bp. CCT is cut less efficiently than CCG and CCA. Some of the CCT sites are not cleaved. (Based on NEB observations.)

Product Source

An E. coli strain that expresses a fusion of Mxe GyrA intein, chitin-binding domain and a truncated form of the Nt.CviPII nicking endonuclease gene from Chlorella virus NYs-1.
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 of pUC19 DNA resulting in a stable pattern of fragments between 25 and 200 bp in 1 hour at 37°C in a total reaction volume of 50 µl.

Reaction Conditions

1X rCutSmart™ Buffer
Incubate at 37°C

1X rCutSmart™ Buffer
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
100 µg/ml Recombinant Albumin
(pH 7.9 @ 25°C)

Activity in NEBuffers

NEBuffer™ r1.1: 10%
NEBuffer™ r2.1: 100%
NEBuffer™ r3.1: 25%
rCutSmart™ Buffer: 100%

Diluent Compatibility

Storage Buffer

20 mM Tris-HCl
100 mM NaCl
50% Glycerol
pH 8 @ 25°C

Heat Inactivation

65°C for 20 minutes

Methylation Sensitivity

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

Product Notes

  1. Tests have suggested that an exonuclease activity is an inherent part of the enzyme. Thus a one hour incubation time is thought to be the optimal time for most procedures. To run on an electrophoresis gel, add loading dye to a final concentration of 0.4% SDS.
  2. Based on the stability of the enzyme in the reaction, incubations longer than 1 hr will not result in improved digestion, unless additional enzyme is added. Please refer to Restriction endonuclease survival in a reaction for more information regarding this topic.
  3. Nt.CviPII is a nicking endonuclease that cleaves only one strand of DNA on a double-stranded DNA substrate. The final product on pUC19 is an array of bands from 25 to 200 bp. CCT is cut less efficiently than CCG and CCA. Some of the CCT sites are not cleaved. (Based on NEB observations.)
  4. Blocked by CpG 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. Open Chromatin Labeling and NicE-seq library construction using Nt.CviPII
  4. Double Digest Protocol with Standard Restriction Enzymes

Usage & Guidelines

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. I tested your restriction enzyme on the substrate DNA recommended by NEB, and it appears to be active, however it does not digest my DNA. What could be the reason?
  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

Tech Tips

Blocked by CpG methylation Enzyme has inherent exonuclease activity

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 [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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