Product Class: Restriction Endonuclease

EcoRI
NEBU cloned at NEB recombinant time saver dil_C 37 65 Heat CpG

EcoRI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10227213. Learn more.

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^AATT_C
Isoschizomers | Single Letter Code

Product Introduction

Catalog # Size Concentration
R0101S 10000 units 20000 units/ml
R0101T 10000 units 100000 units/ml
R0101L 50000 units 20000 units/ml
R0101M 50000 units 100000 units/ml

Featured Videos

View Video Library
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    Reduce Star Activity with High-Fidelity Restriction Enzymes

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    TIME-SAVER™ Protocol for Restriction Enzyme Digests

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    NEB® TV Ep. 15 – Applications of Restriction Enzymes

  • cuttingclose_video_thumb

    Restriction Enzyme Digest Protocol: Cutting Close to DNA End

  • dnasmear_video_thumb

    Restriction Enzyme Digestion Problem: DNA Smear on Agarose Gel

  • EnzymeNotCutting_video_thumb

    Why is My Restriction Enzyme Not Cutting DNA?

  • TooManyBands_video_thumb

    Restriction Enzyme Digest Problem: Too Many DNA Bands

  • nebcloner_dd_demo_thumb

    Double Digestion with NEBcloner

Product Information

Description

EcoRI has a High Fidelity version EcoRI-HF® (NEB #R3101).

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 EcoRI gene from E. coli RY13 (R.N. Yoshimori).
This product is related to the following categories:
Restriction Endonucleases C G,
Time-Saver Qualified Restriction Enzymes,
This product can be used in the following applications:
Fast Cloning: Accelerate your cloning workflows with reagents from NEB,
Restriction Enzyme Digestion

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

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

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™ EcoRI/SspI
Incubate at 37°C

1X NEBuffer™ EcoRI/SspI
100 mM Tris-HCl
50 mM NaCl
10 mM MgCl2
0.025% Triton® X-100
(pH 7.5 @ 25°C)

Activity in NEBuffers

NEBuffer™ r1.1: 25%
NEBuffer™ r2.1: 100%
NEBuffer™ r3.1: 50%
rCutSmart™ Buffer: 50%

Diluent Compatibility

Storage Buffer

10 mM KPO4
300 mM NaCl
0.1 mM EDTA
200 µg/ml Recombinant Albumin
50% Glycerol
1 mM DTT
0.15% Triton® X-100
pH 7 @ 25°C

Heat Inactivation

65°C for 20 minutes

Methylation Sensitivity

dam methylation: Not Sensitive
dcm methylation: Not Sensitive
CpG Methylation: Blocked by Some Combinations of Overlapping

Isoschizomers

EcoRI-HF

Product Notes

  1. May exhibit star activity in NEBuffer r2.1 or rCutSmart Buffer.
  2. This enzyme has shown to have lower activity on some supercoiled plasmids, with more than 1 unit required to digest 1 μg plasmid DNA. For complete digestion of 1 μg of plasmid DNA please follow our recommended digestion protocol..
  3. Blocked by some combinations of overlapping CpG methylation.

Protocols, Manuals & Usage

Protocols

  1. Optimizing Restriction Endonuclease Reactions
  2. Restriction Digest Protocol
  3. Double Digest Protocol with Standard Restriction Enzymes

Usage & Guidelines

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. What is Star Activity and how can it be avoided?
  2. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
  3. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?
  4. Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?

Troubleshooting

Quality, Safety & Legal

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specification Change Notifications

EcoRI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10227213. All subsequent (higher number) lots will contain rAlbumin. For additional information, please visit at NEB Restriction Enzyme formulations with Recombinant Albumin (rAlbumin).

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email [email protected].

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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