Product Class: Restriction Endonuclease

BsiWI-HF®
neb31 cloned at NEB recombinant engineered time saver dil_B 37 80 Heat CpG

BsiWI-HF has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10124088. 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.

NEB high fidelity restriction endonuclease that recognizes the sequence C^GTACG
Isoschizomers | Single Letter Code

Product Introduction

Having supplied restriction enzymes to the research community for over 45 years, NEB has earned the reputation of being the leader in enzyme technologies.  Working continuously to be worthy of that distinction, NEB strives to develop enzyme of the highest purity and unparalleled quality. 

  • Time-Saver™ qualified for digestion in 5-15 minutes
  • 100% activity in rCutSmart™ Buffer (over 210 enzymes are available in the same buffer) allowing for easier double digests
  • Restriction Enzyme Cut Site: C/GTACG
Catalog # Size Concentration
R3553S 300.0 units 20000 units/ml
R3553L 1500.0 units 20000 units/ml

Featured Videos

View Video Library
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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 Digest Problem: Too Many DNA Bands

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    Double Digestion with NEBcloner

Product Information

Description

Product Source

An E. coli strain that carries the cloned and modified BsiWI gene from Bacillus species (D. Clark).
This product is related to the following categories:
High-Fidelity (HF®) Restriction Endonucleases Products,
Restriction Endonucleases B Products,
Time-Saver Qualified Restriction Enzymes Products,
This product can be used in the following applications:
Restriction Enzyme Digestion,
Fast Cloning: Accelerate your cloning workflows with reagents from NEB

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to digest 1 μg of ΦX174 DNA in 1 hour at 37°C in a total reaction volume of 50 μl.

Reaction Conditions

1X rCutSmart™ Buffer
Incubate at 37°C

1X 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: 50%
NEBuffer™ r2.1: 100%
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°C

Heat Inactivation

80°C for 20 minutes

Methylation Sensitivity

dam methylation: Not Sensitive
dcm methylation: Not Sensitive
CpG Methylation: Blocked

Isoschizomers

BsiWI
Pfl23II
PspLI
SplI+

Product Notes

  1. BsiWI-HF has the same specificity as BsiWI (NEB #R0553), but it has been engineered for reduced star activity. 
  2. Blocked by CpG methylation.

Protocols, Manuals & Usage

Protocols

  1. Optimizing Restriction Endonuclease Reactions
  2. Protocol for Direct Digestion of gDNA during droplet digital PCR (ddPCR)
  3. Restriction Digest Protocol
  4. Protocol for Direct Digestion of gDNA during droplet digital PCR (ddPCR)
  5. Double Digest Protocol with Standard Restriction Enzymes

Usage & Guidelines

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. Is there a vector with a unique BsiWI-HF site?
  2. What does the HF® refer to following the name of a restriction enzyme?
  3. How can this enzyme be inactivated?
  4. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
  5. Can Gel Loading Dye, Purple 6X (B7024) be stored in cold temperatures?
  6. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?
  7. Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
  8. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?
  9. Can Gel Loading Dye, Purple 6X (B7024) be stored in cold temperatures?

Troubleshooting

Quality, Safety & Legal

Quality Assurance Statement

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.

Specification Change Notifications

BsiWI-HF has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10124088. All subsequent (higher number) lots will contain rAlbumin. For additional information, please visit at NEB Restriction Enzyme formulations with Recombinant Albumin (rAlbumin).

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]

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]

Safety DataSheets

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 [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.

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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