BstEII

We are excited to announce that we are in the process of switching all reaction buffers to be BSA-free. Beginning April 2021, we will be gradually transitioning to buffers containing Recombinant Albumin (rAlbumin) for restriction enzymes and some DNA modifying enzymes. All information on the website has been updated to reflect this change. Find more details at www.neb.com/BSA-free.

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Having supplied restriction enzymes to the research community for over 45 years, NEB has earned the reputation of being the leader in enzyme technologies. Working continuously to be worthy of that distinction, NEB strives to develop enzyme of the highest purity and unparalleled quality.

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

Catalog #	Concentration	Size	List Price	Your Price
R0162S	10,000 units/ml	2,000 units	$64.00	Sign In Sign In
R0162L	10,000 units/ml	10,000 units	$262.00	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).

Reagents Supplied

The following reagents are supplied with this product:

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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

Product Categories:: Restriction Endonucleases B,; time-saver-qualified-restriction-enzymes Products

Applications:: Fast Cloning: Accelerate your cloning workflows with reagents from NEB,; 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 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
Materials Sold Separately
- NEBuffer™ r3.1
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%