BbsI-HF®
Product information| Code | Name | Size | Quantity | Price | |
|---|---|---|---|---|---|
R3539S |
BbsI-HF |
300 units ( 20000 units/ml ) | - | Unavailable in your region | |
R3539M |
BbsI-HF, conc. |
1000 units ( 50000 units/ml ) | - | Unavailable in your region | |
R3539L |
BbsI-HF |
1500 units ( 20000 units/ml ) | - | Unavailable in your region |
BbsI-HF®
Learn about Ligase Fidelity and Push the Limits of Golden Gate Assembly (50+ fragments).
BbsI-HF has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10119525. Learn more.
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.
High-Fidelity (HF®) restriction enzymes have the same specificity as native enzymes, but have been engineered for significantly reduced star activity and performance in a single buffer (rCutSmart™ Buffer). All HF-restriction enzymes come with Gel Loading Dye, Purple (6X). Enjoy the enhanced performance and added value of our engineered enzymes at the same price as the native enzyme:
- Engineered for improved performance
- 100% activity in rCutSmart Buffer
- Used in Golden Gate Assembly
- sgRNA Template Construction for CRISPR/Cas Genome Editing
- Type IIS restriction enzymes recognize asymmetric DNA sequences and cleave outside of their recognition sequence
- Time-Saver™ qualified for digestion in 5-15 minutes
- Reduced star activity
- Supplied with 1 vial of Gel Loading Dye, Purple (6X)
- Restriction Enzyme Cut Site: GAAGAC(2/6)
- Can be used with NEBridge® Ligase Master Mix for Golden Gate Assembly
Featured Video
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Product Information
Product Source
An E. coli strain that carries the cloned and modified BbsI gene from Bacillus brevis.- This product is related to the following categories:
- Restriction Endonucleases B,
- High-Fidelity (HF®) Restriction Endonucleases,
- Time-Saver Qualified Restriction Enzymes
- This product can be used in the following applications:
- High-throughput cloning and automation solutions,
- NEBridge® Golden Gate Assembly ,
- Restriction Enzyme Digestion
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Reagents Supplied
The following reagents are supplied with this product:
NEB # Component Name Component # Stored at (°C) Amount Concentration -
R3539S -20 BbsI-HF® R3539SVIAL -20 1 x 0.015 ml 20,000 units/ml rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X Gel Loading Dye, Purple (6X) B7024AVIAL 25 1 x 0.5 ml 6 X
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R3539L -20 BbsI-HF® R3539LVIAL -20 1 x 0.075 ml 20,000 units/ml rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X Gel Loading Dye, Purple (6X) B7024AVIAL 25 1 x 0.5 ml 6 X
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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 37°C in a total reaction volume of 50 µl.Reaction Conditions
1X rCutSmart™ Buffer
Incubate at 37°C1X 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: 10%
NEBuffer™ r3.1: 10%
rCutSmart™ Buffer: 100%
Diluent Compatibility
Storage Buffer
10 mM Tris-HCl
300 mM NaCl
1 mM DTT
0.1 mM EDTA
500 µg/ml Recombinant Albumin
50% Glycerol
pH 7.4 @ 25°CHeat Inactivation
65°C for 20 minutesMethylation Sensitivity
dam methylation: Not Sensitive
dcm methylation: Not Sensitive
CpG Methylation: Not Sensitive
Isoschizomers
BbsI
BbvII+
BpiI
BstV2I
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Related Products
Companion Products
Materials Sold Separately
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Product Notes
- BbsI-HF has the same specificity as BbsI (NEB #R0539), but it has been engineered for reduced star activity.
- BbsI is an isoschizomer of BbvII.
- Not sensitive to CpG, dcm, or dam methylation.
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Protocols, Manuals & Usage
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Protocols
- Optimizing Restriction Endonuclease Reactions
- Protocol for Direct Digestion of gDNA during droplet digital PCR (ddPCR)
- Restriction Digest Protocol
- Protocol for Direct Digestion of gDNA during droplet digital PCR (ddPCR)
- Double Digest Protocol with Standard Restriction Enzymes
- Golden Gate Assembly Protocol for NEBridge® Ligase Master Mix (NEB #M1100)
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Usage Guidelines
- 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
- Digestion of Agarose-Embedded DNA: Info for Specific Enzymes
- Double Digests
- Effects of CpG Methylation on Restriction Enzyme Cleavage
- Expanded “assembly standards” for MoClo, GoldenBraid2.0 and other modular Golden Gate Assembly methods
- 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
- Technical Tips For Optimizing Golden Gate Assembly Reactions
- Traditional Cloning Quick Guide
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Tools & Resources
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Selection Charts
- Alphabetized List of Recognition Sequences
- Compatible Cohesive Ends and Generation of New Restriction Sites
- Dam-Dcm and CpG Methylation
- Enzymes with Nonpalindromic Sequences
- Frequencies of Restriction Sites
- Isoelectric Points (pI) for Restriction Enzymes
- Isoschizomers
- NEB Diluent and Buffer Table
- Time-Saver™ Qualified Enzymes
- Type IIS Restriction Enzymes
- Why Choose Recombinant Enzymes?
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Web Tools
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FAQs & Troubleshooting
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FAQs
- What does the HF® refer to following the name of a restriction enzyme?
- Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
- Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?
- Which restriction enzymes are used in GoldenBraid Assembly?
- Which restriction enzymes are used in Golden Gate Assembly?
- Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
- Does NEB offer any BSA-free and/or animal origin-free restriction enzymes for linearization of plasmids for mRNA vaccine development?
- Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?
- Can Gel Loading Dye, Purple 6X (B7024) be stored in cold temperatures?
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Troubleshooting
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Citations & Technical Literature
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Citations
Product Citation Tool
Additional Citations
- Binder A, Lambert J, Morbitzer R, Popp C, Ott T, Lahaye T, Parniske M (2014) A Modular Plasmid Assembly Kit for Multigene Expression, Gene Silencing and Silencing Rescue in Plants PLoS One; 9(2), e88218. PubMedID: 24551083, DOI: 10.1371/journal.pone.0088218
- Alejandro Sarrion-Perdigones, Marta Vazquez-Vilar, Jorge Palací, Bas Castelijns, Javier Forment, Peio Ziarsolo, José Blanca, Antonio Granell, Diego Orzaez (2013) GoldenBraid 2.0: A Comprehensive DNA Assembly Framework for Plant Synthetic Biology Plant Physiol; 162(3), 1618-31. PubMedID: 23669743
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Quality, Safety & Legal
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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
BbsI-HF has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10119525. All subsequent (higher number) lots will contain rAlbumin. For additional information, please visit at NEB Restriction Enzyme formulations with Recombinant Albumin (rAlbumin)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]- R3539S_L_v1_0021612
- R3539S_L_v1_0021701
- R3539S_L_v1_0021609
- R3539S_L_v1_0031703
- R3539S_L_v1_0031705
- R3539S_L_v1_0031709
- R3539S_L_v1_0031804
- R3539S_v1_10013145
- R3539L_v1_10021240
- R3539S_v1_10021239
- R3539S_v1_10031345
- R3539L_v1_10032478
- R3539S_v1_10032476
- R3539S_v1_10037247
- R3539S_v1_10043725
- R3539S_v1_10046382
- R3539L_v1_10047893
- R3539S_v1_10050208
- R3539S_v1_10054974
- R3539L_v1_10056118
- R3539S_v1_10059017
- R3539L_v1_10062015
- R3539S_v1_10063217
- R3539L_v1_10066338
- R3539S_v1_10067980
- R3539L_v1_10071225
- R3539S_v1_10073669
- R3539S_v1_10078550
- R3539S_v1_10079947
- R3539S_v1_10082178
- R3539L_v1_10081999
- R3539S_v1_10086402
- R3539L_v1_10091127
- R3539S_v1_10091114
- R3539S_v1_10089306
- R3539L_v1_10093083
- R3539S_v1_10093081
- R3539L_v1_10101565
- R3539S_v1_10101089
- R3539S_v1_10104551
- R3539L_v1_10107770
- R3539S_v1_10108650
- R3539L_v1_10111180
- R3539S_v1_10114599
- R3539L_v1_10117511
- R3539S_v1_10121029
- R3539L_v2_10119527
- R3539M_v1_10122550
- R3539S_v2_10119525
- R3539S_v2_10129476
- R3539L_v2_10133656
- R3539L_v2_10137679
- R3539S_v2_10135664
- R3539M_v1_10140733
- R3539S_v2_10140844
- R3539L_v2_10140772
- R3539M_v1_10147154
- R3539S_v2_10148322
- R3539L_v2_10149478
- R3539M_v2_10152593
- R3539S_v3_10151471
- R3539L_v3_10152590
- R3539S_v3_10152595
- R3539L_v3_10156939
- R3539S_v3_10159229
- R3539M_v2_10164573
- R3539L_v3_10164024
- R3539S_v3_10167757
- R3539M_v2_10171921
- R3539L_v3_10171914
- R3539S_v3_10171917
- R3539L_v3_10177513
- R3539M_v2_10181310
- R3539M_v2_10187563
- R3539L_v3_10187561
- R3539M_v2_10194873
- R3539S_v3_10191110
- R3539S_v3_10200639
- R3539L_v3_10203018
- R3539M_v2_10205857
- R3539L_v3_10207266
- R3539M_v2_10212855
- R3539S_v3_10207265
- R3539L_v3_10212883
- R3539S_v3_10225332
- R3539S_v3_10217115
- R3539S_v3_10226182
- R3539L_v3_10229220
- R3539M_v2_10231306
- R3539S_v3_10233791
- R3539S_v3_10248424
- R3539L_v3_10249656
- R3539S_v3_10249657
- R3539M_v2_10258492
- R3539S_v3_10261681
- R3539L_v3_10261692
- R3539M_v2_10266146
- R3539S_v3_10279361
- R3539M_v2_10281434
- R3539L_v3_10279364
- R3539S_v3_10283142
- R3539L_v3_10287877
- R3539S_v3_10291112
- R3539L_v3_10302712
- R3539S_v3_10300599
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Safety Data Sheets
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 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.
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Featured Videos
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Golden Gate Assembly Workflow
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Golden Gate Assembly Domestication Tutorial
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Listen to DAD (Data-optimized Assembly Design) when constructing high-complexity Golden Gate Assembly targets
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Golden Gate Assembly Tool Tutorial
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Restriction Enzymes in Golden Gate Assembly
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Reduce Star Activity with High-Fidelity Restriction Enzymes
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TIME-SAVER™ Protocol for Restriction Enzyme Digests
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NEB® TV Ep. 15 – Applications of Restriction Enzymes
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Restriction Enzyme Digest Protocol: Cutting Close to DNA End
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Restriction Enzyme Digestion Problem: DNA Smear on Agarose Gel
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Why is My Restriction Enzyme Not Cutting DNA?
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Restriction Enzyme Digest Problem: Too Many DNA Bands
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Double Digestion with NEBcloner
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Function and Application of Type IIS Restriction Enzymes
Other Products You May Be Interested In
The supporting documents available for this product can be downloaded below.
Protocol
Restriction Digest Protocol
Protocol
Restriction Digest Protocol
Protocol
Restriction Digest Protocol