Product Class: Restriction Endonuclease

recombinant time saver dil_B 37 65 Heat dam Epi


Product Introduction

Having supplied restriction enzymes to the research community for over 40 years, NEB has earned the reputation of being the leader in enzyme technologies.  Working continuously to be worthy of that distinction, NEB strives to develop enzyme of the highest purity and unparalleled quality. 

Bioz Badge Exists : True
Catalog # Size Concentration
R0543S 1,000 units 10,000 units/ml
R0543T 1,000 units 50,000 units/ml
R0543L 5,000 units 10,000 units/ml
R0543M 5,000 units 50,000 units/ml

Product Information


Product Source

An E. coli strain that carries the DpnII gene from Diplococcus pneumoniae G41 (S. Lacks).

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • R0543S     -20    
  • R0543T     -20    
  • R0543L     -20    
  • R0543M     -20    
Product Categories:
Methylation Sensitive Restriction Enzymes for Epigenetics Products,
Restriction Enzymes for Epigenetic Analysis,
Epigenetic Analysis
Restriction Endonucleases C G
time-saver-qualified-restriction-enzymes Products
Restriction Enzymes for Epigenetics,
Fast Cloning: Accelerate your cloning workflows with reagents from NEB,
Restriction Enzyme Digestion

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to digest 1 µg of λ DNA (dam-) in 1 hour at 37°C in a total reaction volume of 50 µl.

Activity in NEBuffers

NEBuffer™ 1.1: 25%
NEBuffer™ 2.1: 25%
NEBuffer™ 3.1: 100%
CutSmart® Buffer: 25%

Diluent Compatibility

Storage Conditions

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

Heat Inactivation

65°C for 20 min

Methylation Sensitivity

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

Product Notes

  1. DpnII and Sau3AI are isoschizomers of MboI.
  2. Cleaves to leave a 5´ GATC extension which can be efficiently ligated to DNA fragments generated by BamHI, BclI, BglII, MboI, Sau3AI and BstYI.
  3. This enzyme is blocked by dam methylation. More information can be found at Dam-Dcm and CpG Methylation.
  4. Will exhibit star activity in NEBuffer 3.1. We recommend the use of NEB DpnII Unique Buffer.

Protocols, Manuals & Usage


  1. Optimizing Restriction Endonuclease Reactions
  2. Double Digest Protocol with Standard Restriction Enzymes
  3. Protocol for Direct Digestion of gDNA during droplet digital PCR (ddPCR)
  4. Restriction Digest Protocol
  5. Protocol for Digestion Prior to droplet digital PCR (ddPCR)

Usage & Guidelines

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting


  1. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
  2. Why is my Restriction Enzyme not cutting DNA?
  3. Why do I see a DNA smear on an agarose gel after a restriction digest?
  4. Why do I see additional DNA bands on my gel after a restriction digest?
  5. How many nucleotides do I have to add adjacent to the RE recognition site in order to get efficient cutting?
  6. Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?


Tech Tips

This enzyme will not cut plasmids isolated from most E.coli because it is blocked by dam methylation. For enzyme cleavage to occur, plasmids must be transformed and isolated from a dam-, dcm- strain of E.coli, such as NEB’s dam-/dcm- Competent E.coli (NEB #C2925).

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.


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

This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

For more information about commercial rights, please contact NEB's Global Business Development team at [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.