Product Class: Restriction Endonuclease

BsmBI-v2
neb31 cloned at NEB recombinant engineered time saver dil_B 55 80 Heat CpG

This product replaces BsmBI (NEB #R0580).

Learn about Ligase Fidelity and Push the Limits of Golden Gate Assembly (50+ fragments).

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.


Catalog #R0739

NEB Type IIS restriction endonuclease that recognizes the sequence CGTCTCN^NNNN and is optimized for Golden Gate Assembly
Isoschizomers | Single Letter Code

Product Introduction

 

Product Information

Description

 

Product Source

An E. coli strain that carries the modified cloned BsmBI gene from Bacillus stearothermophilus B61. BsmBI-v2 is an engineered and improved version of BsmBI. (NEB uses the designation "v2" or "HF" in the name to indicate engineered enzymes.)
This product is related to the following categories:
Restriction Endonucleases B Products,
Time-Saver Qualified Restriction Enzymes Products,
This product can be used in the following applications:
High-throughput cloning and automation solutions,
DNA Preparation,
DNA Analysis,
NEBridge® Golden Gate Assembly ,
Restriction Enzyme Digestion,
Restriction Enzyme Digestion

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 55°C in a total reaction volume of 50 µl.

Reaction Conditions

1X NEBuffer™ r3.1
Incubate at 55°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)

Usage Concentration

1X

Activity in NEBuffers

NEBuffer™ r1.1: <10%
NEBuffer™ r2.1: 50%
NEBuffer™ r3.1: 100%
rCutSmart™ Buffer: 25%

Diluent Compatibility

Storage Buffer

10 mM Tris-HCl
300 mM NaCl
1 mM DTT
0.1 mM EDTA
500 µg/ml BSA
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

Activity at Temperature

@37°C: 10%

Product Notes

  1. BsmBI-v2 is an isoschizomer of Esp3I

Protocols, Manuals & Usage

Protocols

  1. Optimizing Restriction Endonuclease Reactions
  2. Restriction Digest Protocol
  3. Double Digest Protocol with Standard Restriction Enzymes
  4. 35-Fragment Golden Gate Assembly using BsmBI-v2 (NEB #E1602)
  5. Golden Gate Assembly Protocol for using NEBridge® Golden Gate Assembly Kit (BsmBI-v2) (NEB #E1602)

Usage & Guidelines

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. Why is there a BsmBI site in LITMUS 38 but not LITMUS 39?
  2. Which restriction enzymes are used in Golden Gate Assembly?
  3. Which restriction enzymes are used in GoldenBraid Assembly?
  4. When is star activity a concern?
  5. Is Gel Loading Dye, Purple (6X) or Gel Loading Dye, Purple (6X), no SDS compatible with other DNA binding dyes such as SYBR® and GelRed™ during gel electrophoresis?
  6. How many nucleotides do I have to add adjacent to the RE recognition site in order to get efficient cutting?
  7. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?
  8. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?
  9. What are the differences between BsmBI-v2 and its isoschizomer, Esp3I?

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.

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 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.

Other Products You May Be Interested In