Home Restriction Endonucleases BbsI

BbsI

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

BbsI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10166193. Learn more.

NEB restriction endonuclease that recognizes the sequence GAAGACNN^NNNN_

Isoschizomers | Single Letter Code | Pronunciation:

Time-Saver™ qualified for digestion in 5-15 minutes
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
High Fidelity (HF^®) version available (NEB #R3539) supplied with rCutSmart^™ Buffer
Supplied with 1 vial of Gel Loading Dye, Purple (6X)
Restriction Enzyme Cut Site: GAAGAC(2/6)

Catalog #	Cat #	Concentration	Conc	Size	List Price
R0539S	10,000 units/ml	300 units	$83.00	$83.00	Sign In Sign In
R0539L	10,000 units/ml	1,500 units	$343.00	$343.00	Sign In Sign In

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Product Information

BbsI has a High Fidelity version BbsI-HF^® (NEB #R3539).

High Fidelity (HF) Restriction Enzymes have 100% activity in rCutSmart 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 minutes 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.

For details on NEB’s quality controls for restriction endonucleases, visit our Restriction Enzyme Quality page.

Product Source

An E. coli strain that carries the cloned BbsI gene from Bacillus brevis.

This product is related to the following categories:: Restriction Endonucleases B,; Time-Saver Qualified Restriction Enzymes

This product can be used in the following applications:: NEBridge® Golden Gate Assembly ,; Fast Cloning: Accelerate your cloning workflows with reagents from NEB,; Restriction Enzyme Digestion

Reagents Supplied

The following reagents are supplied with this product:

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

R0539S

-20

BbsI	R0539SVIAL	-20	1 x 0.03 ml	10,000 units/ml
NEBuffer™ r2.1	B6002SVIAL	-20	1 x 1.25 ml	10 X
Gel Loading Dye, Purple (6X)	B7024AVIAL	25	1 x 0.5 ml	6 X

R0539L

-20

BbsI	R0539LVIAL	-20	1 x 0.15 ml	10,000 units/ml
NEBuffer™ r2.1	B6002SVIAL	-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 NEBuffer™ r2.1
Incubate at 37°C

1X NEBuffer™ r2.1
50 mM NaCl
10 mM Tris-HCl
10 mM MgCl₂
100 µg/ml Recombinant Albumin
(pH 7.9 @ 25°C)

Activity in NEBuffers
NEBuffer™ r1.1: 100%
NEBuffer™ r2.1: 100%
NEBuffer™ r3.1: 25%
rCutSmart™ Buffer: 75%

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

Isoschizomers

BbsI
BbsI-HF
BbvII+
BpiI
BstV2I
Related Products
Companion Products
- BbsI-HF^®
Materials Sold Separately
- Gel Loading Dye, Purple (6X)
Product Notes
1. The enzyme retains full activity after at least 10 free-thaw cycles; the enzyme can be aliquoted and stored at –80°C.
2. BbsI is an isoschizomer of BbvII.
3. Not sensitive to CpG, dcm, or dam methylation.