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Home Restriction Endonucleases Products BclI-HF

BclI-HF

Bcl-I-cutsite_1

Isoschizomers | Single Letter Code | Pronunciation:

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 (CutSmart^® 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 CutSmart Buffer
Time-Saver™ qualified for digestion in 5-15 minutes
Reduced star activity
Supplied with 1 vial of Gel Loading Dye, Purple (6X)

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 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 cloned and modified BclI gene from Bacillus caldolyticus (A. Atkinson).
Reagents Supplied

The following reagents are supplied with this product:

Store at (°C) Concentration

Gel Loading Dye, Purple (6X) 25 6 X

CutSmart® Buffer -20 10 X

Product Categories:

High-Fidelity (HF®) Restriction Endonucleases Products,

Restriction Endonucleases B,

time-saver-qualified-restriction-enzymes Products

Applications:

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.
  Reaction Conditions
  
  1X CutSmart® Buffer
  Incubate at 37°C
  
  1X CutSmart® Buffer
  50 mM Potassium Acetate
  20 mM Tris-acetate
  10 mM Magnesium Acetate
  100 µg/ml BSA
  (pH 7.9 @ 25°C)
  
  Activity in NEBuffers
  CutSmart® Buffer: 100%
  NEBuffer™ 1.1: 100%
  NEBuffer™ 2.1: 100%
  NEBuffer™ 3.1: 10%
  
  Diluent Compatibility
  
  Diluent B
  
  Storage Temperature
  -20°C
  Storage Conditions
  
  10 mM Tris-HCl
  300 mM NaCl
  1 mM DTT
  0.1 mM EDTA
  200 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
- Related Products
  Companion Products
  
  Gel Loading Dye, Purple (6X)
  
  dam^-/dcm^- Competent E. coli
  
  Nuclease-free Water
  
  BclI
  
  Monarch® Plasmid Miniprep Kit
  
  Monarch® DNA Gel Extraction Kit
  
  Monarch® PCR & DNA Cleanup Kit (5 μg)
  
  Materials Sold Separately
  
  CutSmart® Buffer
- Product Notes
  BclI-HF has the same specificity as BclI (NEB #R0160), but it has been engineered for reduced star activity.
  
  Cleaves to leave a 5´ GATC extension which can be efficiently ligated to DNA fragments generated by BamHI, BglII, MboI, Sau3AI and BstYI.
  
  This enzyme is blocked by dam methylation. More information can be found at Dam-Dcm and CpG Methylation.
Protocols, Manuals & Usage
- Protocols
  Optimizing Restriction Endonuclease Reactions
  
  Double Digest Protocol with Standard Restriction Enzymes
  
  Restriction Digest Protocol
- Usage Guidelines
  Activity of Restriction Enzymes in PCR Buffers
  
  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
  
  General Molecular Biology Data
  
  Getting Started with Molecular Cloning: Simple Tips to Improve your Cloning Efficiency
  
  Heat Inactivation
  
  Making Unmethylated (Dam- Dcm-) DNA
  
  NEBuffer Activity/Performance Chart with Restriction Enzymes
  
  Optimizing Restriction Endonuclease Reactions
  
  Reduced Star Activities of HF® Enzymes
  
  Restriction Endonucleases - Survival in a Reaction
  
  Restriction Enzyme Diluent Buffer Compatibility
  
  Restriction Enzyme Tips
  
  Single Letter Codes
  
  Six Tips for a Perfect Gel Extraction
  
  Star Activity
  
  The Genetic Code
  
  Traditional Cloning Quick Guide
Tools & Resources
- Selection Charts
  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
  
  Enzyme Finder - Sequence > 6 Bases
  
  Frequencies of Restriction Sites
  
  Isoschizomers
  
  Recleavable Filled-in 5' Overhangs
  
  Time-Saver™ Qualified Enzymes
  
  Why Choose Recombinant Enzymes?
- Web Tools
  Competitor Cross-Reference Tool
  
  DNA Sequences and Maps Tool
  
  Enzyme Finder
  
  NEBcutter V2.0
  
  NEBioCalculator
  
  REBASE
FAQs & Troubleshooting
- FAQs
  What does HF® refer to following the name of a restriction enzyme?
  
  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?
  
  Can Gel Loading Dye, Purple 6X (B7024) be stored in cold temperatures?
  
  Why is my Restriction Enzyme not cutting DNA?
  
  Why do I see additional DNA bands on my gel after a restriction digest?
  
  Why do I see a DNA smear on an agarose 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?
  
  Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
- Troubleshooting
  Restriction Enzyme Troubleshooting Guide
- 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 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.
- 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]
  
  R3160S_L_v1_0021705
  
  R3160S_L_v1_0021801
  
  R3160L_v1_10011637
  
  R3160S_v1_10011639
  
  R3160L_v1_10019564
  
  R3160S_v1_10017518
  
  R3160S_v1_10028230
  
  R3160L_v1_10042363
  
  R3160S_v1_10042361
  
  R3160L_v1_10060988
  
  R3160S_v1_10062206
- Safety Data Sheets
  The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
  
  BclI-HF
  
  Gel Loading Dye, Purple (6X)
  
  CutSmart® Buffer
- 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).
  
  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.
  
  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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