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Home Restriction Endonucleases Products BstNI

BstNI

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 CC^W_GG

Isoschizomers | Single Letter Code | Pronunciation:

Time-Saver™ qualified for digestion in 5-15 minutes
Restriction Enzyme Cut Site: CC/WGG

Catalog #	Concentration	Size	List Price	Your Price
R0168S	10,000 units/ml	3,000 units	$67.00	Sign In Sign In
R0168L	10,000 units/ml	15,000 units	$273.00	Sign In Sign In

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Product Information

Product Source

An E. coli strain that carries the cloned BstNI gene from Bacillus stearothermophilus N (D. Comb)

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

R0168S -20

BstNI R0168SVIAL -20 1 x 0.3 ml 10,000 units/ml

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

R0168L -20

BstNI R0168LVIAL -20 2 x 0.75 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: 100%
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

Isoschizomers

AjnI
BciT130I
BseBI
Bst2UI
EcoRII
MvaI
Psp6I
PspGI

Activity at Temperature
@37°C: 25%
Related Products
Companion Products
Product Notes
1. BstNI is an isoschizomer of EcoRII but cuts DNA at a different location. (EcoRII cuts before the two C residues).
2. BstNI produces DNA fragments that have a single-base 5´ extension which are more difficult to ligate than blunt-ended fragments. More efficient ligation can be achieved by using the Quick Ligation Kit (NEB #M2200).
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. For enzymes that cannot be heat-inactivated, we recommend using a column for cleanup (such as the Monarch® PCR & DNA Cleanup Kit), or running the reaction on an agarose gel and then extracting the DNA (we recommend Monarch Gel Extraction Kit), or performing a phenol/chloroform extraction.
5. Not sensitive to CpG, dcm, or dam methylation.