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 worth of that distinction, NEB strives to develop enzyme of the highest purity and unparalleled quality.
Restriction Enzymes in Golden Gate Assembly
Golden Gate Assembly Workflow
Reduce Star Activity with High-Fidelity Restriction Enzymes
Time-Saver Protocol for Restriction Enzyme Digests
NEB TV Episode 15
CutSmart Restriction Enzyme Buffer
Restriction Enzyme Digest Protocol: Cutting Close to DNA End
Restriction Enzyme Digestion Problem: DNA Smear on Agarose Gel
Why is My Restriction Enzyme Not Cutting DNA?
Product SourceAn E. coli strain that carries the cloned BspQI gene from Bacillus sphaericus (X.S. Pan)
The following reagents are supplied with this product:
|Store at (°C)||Concentration|
|NEBuffer™ 3.1||-20||10 X|
Properties and Usage
Unit DefinitionOne unit is defined as the amount of enzyme required to digest 1 µg of λ DNA in 1 hour at 50°C in a total reaction volume of 50 µl.
1X NEBuffer™ 3.1
Incubate at 50°C
1X NEBuffer™ 3.1
100 mM NaCl
50 mM Tris-HCl
10 mM MgCl2
100 μg/ml BSA
(pH 7.9 @ 25°C)
Activity in NEBuffersCutSmart® Buffer: 100%
NEBuffer 1.1: 100%
NEBuffer 2.1: 100%
NEBuffer 3.1: 100%
20 mM Tris-HCl
500 mM KCl
1 mM DTT
0.1 mM EDTA
500 μg/ml BSA
0.1% Triton® X-100
pH 7.0 @ 25°C
Heat Inactivation80°C for 20 min
dam methylation: Not Sensitive
dcm methylation: Not Sensitive
CpG Methylation: Not Sensitive
Activity at Temperature@37°C: 10%
- 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.
Faqs & Tech Tips
- What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
- Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
- How can I access the old NEBuffer Activity Chart?
- Why is my Restriction Enzyme not cutting DNA?
- Why do I see a DNA smear on an agarose gel after a restriction digest?
- Why do I see additional DNA bands on my gel after a restriction digest?
- How many nucleotides do I have to add adjacent to the RE recognition site in order to get efficient cutting?
Protocols & Manuals
Other Tools & Resources
- Alphabetized List of Recognition Specificities
- Buffer and Diluent Formulation Table
- Compatible Cohesive Ends and Generation of New Restriction Sites
- Cross Index of Recognition Sequences
- Dam-Dcm and CpG Methylation
- Enzymes with Nonpalindromic Sequences
- Frequencies of Restriction Sites
- Time-Saver™ Qualified Enzymes
- Type IIS Restriction Enzymes
- Why Choose Recombinant Enzymes?
Usage Guildelines & Tips
- Activity at 37°C for Restriction Enzymes with Alternate Incubation Temperatures
- Activity of Restriction Enzymes in PCR Buffers
- Cleavage Close to the End of DNA Fragments
- Cleavage of Supercoiled DNA
- Digestion of Agarose-Embedded DNA: Info for Specific Enzymes
- Double Digests
- Heat Inactivation
- Megabase Mapping
- NEBuffer Activity/Performance Chart with Restriction Enzymes
- Optimizing Restriction Endonuclease Reactions
- Restriction Endonucleases - Survival in a Reaction
- Restriction Enzyme Diluent Buffer Compatibility
- Restriction Enzyme Tips
- Single Letter Codes
- Star Activity
- Traditional Cloning Quick Guide
Troubleshooting GuidesRestriction Enzyme Troubleshooting Guide
Quality & Safety
Quality Assurance StatementQuality 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.
SpecificationsThe 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 AnalysisThe Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality control's for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Safety DataSheetsThe following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
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@example.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.