BsaI-HF^®v2

BsaI-HFv2 has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10156743. Learn more.

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.

BsaI-HFv2 is NEB Type IIS restriction enzyme that has been optimized for Golden Gate Assembly and in protocols requiring DNNEB cutting by BsaI Works well for any protocol requiring DNNEB cutting by BsaI Recommended enzyme for any purpose requiring digestion at the recognition sequence 5-GGTCTC(N1)/(N5)-3

Isoschizomers | Single Letter Code | Pronunciation:

Attention Golden Gate Assembly users: BsaI-HFv2 (NEB #R3733) has been optimized for Golden Gate Assembly*. BsaI-HFv2 also works well for any protocol requiring DNA cutting by BsaI. This is the recommended enzyme for any purpose requiring digestion at the recognition sequence ...5′-GGTCTC(N1)/(N5)-3′...

*The requirements for digestion of DNA during Golden Gate Assembly are more demanding than what is encountered in traditional DNA cloning. For Golden Gate, the enzyme must function well in a buffer that might not be the same buffer as that normally provided for the restriction enzyme, and maintain activity for an extended time at elevated temperatures in a dynamic cutting/re-ligating reaction with competition for substrate binding between the endonuclease and ligase. Extensive testing has demonstrated superior performance of BsaI-HFv2 compared to both BsaI (NEB #R0535) and BsaI-HF (NEB #R3535) in this challenging Golden Gate assembly context where restriction enzyme efficiency and fidelity are critically important.

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
Used in Golden Gate Assembly; maintains high activity levels in T4 DNA Ligase Buffer often used in this application
Type IIS restriction enzymes recognize asymmetric DNA sequences and cleave outside of their recognition sequence
100% activity in rCutSmart Buffer
Time-Saver™ qualified for digestion in 5-15 minutes
Reduced star activity
NEB has developed convenient kits (using BsmBI-v2 and BsaI-HFv2) for performing Golden Gate Assembly.
Restriction Enzyme Cut Site: GGTCTC(1/5)

"I really, really love BsaI-HFv2. We are back to getting phenomenal GGA results again with very little background."

-M.C, Principal Investigator, Davidson College, NC

Catalog #	Concentration	Size	List Price	Your Price
R3733S	20,000 units/ml	1,000 units	$75.00	Sign In Sign In
R3733L	20,000 units/ml	5,000 units	$308.00	Sign In Sign In

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Product Information

Product Source

An E. coli strain that carries the cloned and modified BsaI gene from Bacillus stearothermophilus 6-55 (Z. Chen)

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:: NEBridge® Golden Gate Assembly ,; Restriction Enzyme Digestion,; Fast Cloning: Accelerate your cloning workflows with reagents from NEB,

Reagents Supplied

The following reagents are supplied with this product:

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

R3733S

-20

BsaI-HF^®v2	R3733SVIAL	-20	1 x 0.05 ml	20,000 units/ml
Gel Loading Dye, Purple (6X)	B7024AVIAL	25	1 x 0.5 ml	6 X
rCutSmart™ Buffer	B6004SVIAL	-20	1 x 1.25 ml	10 X

R3733L

-20

BsaI-HF^®v2	R3733LVIAL	-20	1 x 0.25 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

Properties & Usage
Unit Definition
One unit is defined as the amount of enzyme required to digest 1 µg of pXba 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: 100%
NEBuffer™ r2.1: 100%
NEBuffer™ r3.1: 100%
rCutSmart™ Buffer: 100%

Diluent Compatibility
- Diluent B
Storage Buffer

20 mM Tris-HCl
300 mM NaCl
0.1 mM TCEP
200 µg/ml Recombinant Albumin
50% Glycerol
pH 9 @ 25°C

Heat Inactivation
80°C for 20 minutes
Methylation Sensitivity

dam methylation: Not Sensitive
dcm methylation: Impaired by Some Combinations of Overlapping
CpG Methylation: Blocked by Some Combinations of Overlapping
Related Products
Companion Products
Product Notes
1. BsaI-HFv2 has the same specificity as BsaI (NEB #R0535), but it has been engineered for reduced star activity.
2. Impaired by some combinations of overlapping dcm methylation and blocked by some combinations of overlapping CpG methylation.