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BstEII

This product was discontinued on 12/15/2022 and is replaced with the high fidelity version, BstEII-HF™ (NEB #R3162).

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.

Replaced by BstEII-HF® on December 15, 2022

Catalog # R0162 was discontinued on December 20, 2022

NEB restriction endonuclease that recognizes the sequence G^GTNAC_C

Isoschizomers | Single Letter Code | Pronunciation:

Time-Saver™ qualified for digestion in 5-15 minutes
High Fidelity (HF^®) version available (NEB #R3162) supplied with rCutSmart^™ Buffer
Restriction Enzyme Cut Site: G/GTNACC

Product Information
BstEII has a High Fidelity version, BstEII-HF® (NEB #R3162)

High Fidelity (HF) Restriction Enzymes have 100% activity in rCutSmart 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).

This product is related to the following categories:

Discontinued (<3 years)
- Properties & 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™ r3.1
  Incubate at 60°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: 10%
  NEBuffer™ r2.1: 75%
  NEBuffer™ r3.1: 100%
  rCutSmart™ Buffer: 75%
  
  Diluent Compatibility
  
  Diluent A
  
  Storage Buffer
  
  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: 10%
- Related Products
  Companion Products
  
  BstEII-HF®
  
  Monarch® Plasmid Miniprep Kit
  
  Monarch® DNA Gel Extraction Kit
  
  Monarch® PCR & DNA Cleanup Kit (5 μg)
- Product Notes
  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 r2.1 and rCutSmart Buffer.
  
  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.
  
  Not sensitive to CpG, dcm, or dam methylation.
  
  star activity may result from a glycerol concentration of >5%.
Protocols, Manuals & Usage
- Protocols
  Optimizing Restriction Endonuclease Reactions
  
  Restriction Digest Protocol
  
  Double Digest Protocol with Standard Restriction Enzymes
- Usage Guidelines
  Activity at 37°C for Restriction Enzymes with Alternate Incubation Temperatures
  
  Cleavage Close to the End of DNA Fragments
  
  Dam and Dcm Methylases of E. coli
  
  Digestion of Agarose-Embedded DNA: Info for Specific Enzymes
  
  Double Digests
  
  Effects of CpG Methylation on Restriction Enzyme Cleavage
  
  Heat Inactivation
  
  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
Tools & Resources
- Selection Charts
  Alphabetized List of Recognition Sequences
  
  Compatible Cohesive Ends and Generation of New Restriction Sites
  
  Dam-Dcm and CpG Methylation
  
  Enzymes with Multiple Recognition Sequences
  
  Frequencies of Restriction Sites
  
  Interrupted Palindromes
  
  Isoelectric Points (pI) for Restriction Enzymes
  
  Isoschizomers
  
  Time-Saver™ Qualified Enzymes
  
  Why Choose Recombinant Enzymes?
- Web Tools
  Competitor Cross-Reference Tool
  
  DNA Sequences and Maps Tool
  
  Double Digest Finder
  
  Enzyme Finder
  
  NEBcutter™ v3.0
  
  NEBioCalculator®
  
  REBASE®
FAQs & Troubleshooting
- FAQs
  Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
  
  Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
  
  Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?
- Troubleshooting
  Restriction Enzyme Troubleshooting Guide
- 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.
Citations & Technical Literature
- Citations
  
  Product Citation Tool
  
  Powered by Bioz See more details on Bioz
Quality, Safety & Legal
- 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 & Change Notifications
  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
  
  R0162S_L_v2
  
  R0162T_M_v2
- 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
  
  R0162S_v1_10031296
  
  R0162L_v1_10044563
  
  R0162M_v1_10043559
  
  R0162S_v1_10049593
  
  R0162L_v1_10052081
  
  R0162M_v1_10063036
  
  R0162L_v1_10065275
  
  R0162S_v1_10065273
  
  R0162L_v1_10082797
  
  R0162S_v1_10086406
  
  R0162L_v1_10091787
  
  R0162S_v1_10093538
  
  R0162L_v1_10101984
  
  R0162L_v1_10116393
  
  R0162S_v1_10116394
  
  R0162S_v2_10139468
  
  R0162L_v2_10143505
  
  R0162S_v2_10152004
  
  R0162L_v2_10157929
- 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
- 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 busdev@neb.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.
  
  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.
  Trademarks

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