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Esp3I

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.

NEB Type IIS restriction endonuclease that recognizes the sequence CGTCTCN^NNNN_

Isoschizomers | Single Letter Code | Pronunciation:

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 enzymes of the highest purity and unparalleled quality.

Time-Saver™ qualified for digestion in 5-15 minutes
Isoschizomer of BsmBI-v2
Used in Golden Gate Assembly
Type IIS restriction enzymes recognize asymmetric DNA sequences and cleave outside of their recognition sequence
Supplied with 1 vial of Gel Loading Dye, Purple (6X)
NEB has developed convenient kits (using BsmBI-v2 and BsaI-HFv2) for performing Golden Gate Assembly.
Restriction Enzyme Cut Site: CGTCTC(1/5)
Can be used with NEBridge^® Ligase Master Mix for Golden Gate Assembly

Catalog #	Concentration	Size	List Price	Your Price
R0734S	10,000 units/ml	300 units	$85.00	Sign In Sign In
R0734L	10,000 units/ml	1,500 units	$392.00	Sign In Sign In

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Product Information

Product Source

An E. coli strain that carries the cloned Esp3I gene from Erwinia species RFL3 (A. Janulaitis).

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

This product can be used in the following applications:: High-throughput cloning and automation solutions,; NEBridge® Golden Gate Assembly ,; 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

R0734S -20

Esp3I R0734SVIAL -20 1 x 0.03 ml 10,000 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

R0734L -20

Esp3I R0734LVIAL -20 1 x 0.15 ml 10,000 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 λ 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: <10%
rCutSmart™ Buffer: 100%

Diluent Compatibility
- Diluent B
Storage Buffer

10 mM Tris-HCl
300 mM NaCl
1 mM DTT
0.1 mM EDTA
500 µg/ml BSA
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: Blocked
Related Products
Companion Products
Materials Sold Separately
- rCutSmart™ Buffer
- Gel Loading Dye, Purple (6X)
Product Notes
1. Esp3I is an isoschizomer of BsmBI-v2
2. This enzyme is blocked by Cpg methylation. More information can be found at "Dam-Dcm and CpG Methylation".