BstEII

This product will be discontinued on 12/15/2022 and is replaced with the high fidelity version, BstEII-HF™ (NEB #R3162).

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 restriction endonuclease that recognizes the sequence G^GTNAC_C

Isoschizomers | Single Letter Code | Pronunciation:

Time-Saver™ qualified for digestion in 5-15 minutes
High Fidelity (HF^®) version available (NEB #R3162) supplied with rCutSmart^™ Buffer
Restriction Enzyme Cut Site: G/GTNACC

Catalog #	Concentration	Size	List Price	Your Price
R0162S	10,000 units/ml	2,000 units	$66.00	Sign In Sign In
R0162L	10,000 units/ml	10,000 units	$270.00	Sign In Sign In

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Product Information

BstEII has a High Fidelity version, BstEII-HF® (NEB #R3162)

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.

Product Source

An E. coli strain that carries the BstEII gene from Bacillus stearothermophilus ET (N. Welker).

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

This product can be used in the following applications:: 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

R0162S -20

BstEII R0162SVIAL -20 1 x 0.2 ml 10,000 units/ml

NEBuffer™ r3.1 B6003SVIAL -20 1 x 1.25 ml 10 X

R0162L -20

BstEII R0162LVIAL -20 1 x 1 ml 10,000 units/ml

NEBuffer™ r3.1 B6003SVIAL -20 1 x 1.25 ml 10 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 60°C in a total reaction volume of 50 µl.
Reaction Conditions

1X NEBuffer™ r3.1
Incubate at 60°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)

Activity in NEBuffers
NEBuffer™ r1.1: 10%
NEBuffer™ r2.1: 75%
NEBuffer™ r3.1: 100%
rCutSmart™ Buffer: 75%

Diluent Compatibility
- Diluent A
Storage Buffer

10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
200 µg/ml BSA
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation
No
Methylation Sensitivity

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

Activity at Temperature
@37°C: 10%
Related Products
Companion Products
Product Notes
1. Of the over 3,000 known restriction endonucleases, BstEII is one of the few that produces extensions of more than 4 bases.
2. May exhibit Star Activity in NEBuffer r2.1 and rCutSmart Buffer.
3. Based on the stability of the enzyme in the reaction, incubations longer than 1 hr will not result in improved digestion, unless additional enzyme is added. Please refer to Restriction endonuclease survival in a reaction for more information regarding this topic.
4. Not sensitive to CpG, dcm, or dam methylation.
5. star activity may result from a glycerol concentration of >5%