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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.

NEB high fidelity restriction endonuclease that recognizes the sequence GAAGAC(N)2^

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 (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

Catalog #	Concentration	Size	List Price	Your Price
R3539S	20,000 units/ml	300 units	$78.00	Sign In Sign In
R3539L	20,000 units/ml	1,500 units	$323.00	Sign In Sign In
R3539M	50,000 units/ml	1,000 units	$162.00	Sign In Sign In

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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 Products,; High-Fidelity (HF®) Restriction Endonucleases Products,; Time-Saver Qualified Restriction Enzymes Products

This product can be used in the following applications:: High-throughput cloning and automation solutions,; NEBridge® Golden Gate Assembly ,; Restriction Enzyme Digestion

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

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

R3539M -20

BbsI-HF^® R3539MVIAL -20 1 x 0.02 ml 50,000 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

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°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: 10%
NEBuffer™ r2.1: 10%
NEBuffer™ r3.1: 10%
rCutSmart™ Buffer: 100%

Diluent Compatibility
- Diluent B
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
65°C for 20 minutes
Methylation Sensitivity

dam methylation: Not Sensitive
dcm methylation: Not Sensitive
CpG Methylation: Not Sensitive
Related Products
Companion Products
Materials Sold Separately
- rCutSmart™ Buffer
Product Notes
1. BbsI-HF has the same specificity as BbsI (NEB #R0539), but it has been engineered for reduced star activity.
2. BbsI is an isoschizomer of BbvII.
3. Not sensitive to CpG, dcm, or dam methylation.