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BstEII

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incubation temp

BstE-II-cutsite_1

Isoschizomers | Single Letter Code

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.

Time-Saver™ qualified for digestion in 5-15 minutes
High Fidelity (HF^®) version available (NEB #R3162) supplied with CutSmart^® Buffer

Featured Videos

View Video Library

Reduce Star Activity with High-Fidelity Restriction Enzymes
Time-Saver Protocol for Restriction Enzyme Digests
NEB TV Ep. 15 – Applications of Restriction Enzymes
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?
Restriction Enzyme Digest Problem: Too Many DNA Bands
Double Digestion with NEBcloner

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Ordering Information

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Product Information
High Fidelity (HF) Restriction Enzymes have 100% activity in CutSmart Buffer; single-buffer simplicity means more straightforward and streamlined sample processing. HF enzymes also exhibit dramatically reduced star activity. HF enzymes are all Time-Saver qualified and can therefore cut substrate DNA in 5-15 minutes with the flexibility to digest overnight without degradation to DNA. Engineered with performance in mind, HF restriction enzymes are fully active under a broader range of conditions, minimizing off-target products, while offering flexibility in experimental design.
Product Source
An E. coli strain that carries the BstEII gene from Bacillus stearothermophilus ET (N. Welker).
Reagents Supplied

The following reagents are supplied with this product:

Store at (°C) Concentration

NEBuffer™ 3.1 -20 10 X

Product Categories:

Restriction Endonucleases B,

time-saver-qualified-restriction-enzymes

Applications:

Fast Cloning: Accelerate your cloning workflows with reagents from NEB,

Restriction Enzyme Digestion
- Properties and Usage
  Unit Definition
  One unit is defined as the amount of enzyme required to digest 1 µg of λ DNA in 1 hour at 60°C in a total reaction volume of 50 µl.
  Reaction Conditions
  
  1X NEBuffer™ 3.1
  Incubate at 60°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 NEBuffers
  CutSmart® Buffer: 75%
  NEBuffer 1.1: 10%
  NEBuffer 2.1: 75%
  NEBuffer 3.1: 100%
  
  Diluent Compatibility
  
  Diluent A
  
  Storage Temperature
  -20°C
  Storage Conditions
  
  10 mM Tris-HCl
  50 mM KCl
  1 mM DTT
  0.1 mM EDTA
  200 μg/ml BSA
  50% Glycerol
  pH 7.4 @ 25°C
  
  Heat Inactivation
  No
  Methylation Sensitivity
  
  dam methylation: Not Sensitive
  dcm methylation: Not Sensitive
  CpG Methylation: Not Sensitive
  
  Activity at Temperature
  @37°C: 50%
- Related Products
  Companion Products
  
  BstEII-HF®
  
  Monarch® Plasmid Miniprep Kit
  
  Monarch® DNA Gel Extraction Kit
  
  Monarch® PCR & DNA Cleanup Kit (5 μg)
  
  Materials Sold Separately
  
  NEBuffer 3.1
- Notes
  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.
  
  Of the over 3,000 known restriction endonucleases, BstEII is one of the few that produces extensions of more than 4 bases.
  
  May exhibit Star Activity in NEBuffer 2.1 and CutSmart Buffer.
  
  Star activity may result from a glycerol concentration of > 5%.
FAQs & Tech Tips
- FAQs
  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?
- Tech Tips
  
  BstEII will exhibit star activity in a low salt buffer.
  
  You will not get additional cutting after incubation at 60C for 1-2hr.
Protocols & Manuals
- Protocols
  Optimizing Restriction Endonuclease Reactions
  
  Double Digest Protocol with Standard Restriction Enzymes
  
  Restriction Digest Protocol
Other Tools & Resources
Quality & Safety
- Quality Control Assays
  
  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]
  
  R0162S_L_v1
  
  R0162T_M_v1
- 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]
  
  R0162S_L_v1_0851309
  
  R0162S_L_v1_0851405
  
  R0162S_L_v1_0841212
  
  R0162S_L_v1_0841304
  
  R0162T_M_v1_0841212
  
  R0162T_M_v1_0841304
  
  R0162T_M_v1_0851405
  
  R0162T_M_v1_0851410
  
  R0162S_L_v1_0851410
  
  R0162S_L_v1_0851504
  
  R0162T_M_v1_0851504
  
  R0162T_M_v1_0851509
  
  R0162S_L_v1_0851510
  
  R0162S_L_v1_0851604
  
  R0162T_M_v1_0851606
  
  R0162S_L_v1_0851610
  
  R0162T_M_v1_0851703
  
  R0162S_L_v1_0851709
  
  R0162T_M_v1_0851712
  
  R0162S_L_v1_0851804
  
  R0162L_v1_10008590
  
  R0162M_v1_10030264
  
  R0162L_v1_10028212
  
  R0162S_v1_10028214
- Safety Data Sheets
  The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
  
  BstEII
  
  NEBuffer™ 3.1
Legal Information
- 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.

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