Product Class: Restriction Endonuclease

EcoRI

icon_cloned icon_recomb icon_timesaver5 icon_NEBU dil_C-grey icon_incTemp37 icon_HI_65 icon_cpg icon_bluewhite
EcoR-I-cutsite_1

Product Introduction

Having supplied restriction enzymes to the research community for over 40 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.

Catalog # Size Concentration
R0101L 50,000 units 20,000 units/ml
R0101M 50,000 units 100,000 units/ml
R0101S 10,000 units 20,000 units/ml
R0101T 10,000 units 100,000 units/ml

Product Information

Description

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

High Fidelity (HF) Restriction Enzymes have 100% activity in CutSmart 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).

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C) Concentration
NEBuffer™ EcoRI -20 10 X
Product Categories:
Restriction Endonucleases: C-G,
Time-Saver™ Qualified Restriction Enzymes
Applications:
Fast Cloning: Accelerate your cloning workflows with reagents from NEB,
Restriction Enzyme Digestion

Properties and 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
Incubate at 37°C

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

Activity in NEBuffers

CutSmart® Buffer: 50%
NEBuffer 1.1: 25%
NEBuffer 2.1: 100%
NEBuffer 3.1: 50%

Diluent Compatibility

Storage Temperature

-20°C

Storage Conditions

10 mM KPO4
300 mM NaCl
0.1 mM EDTA
200 μg/ml BSA
50% Glycerol
1 mM DTT
0.15% Triton® X-100
pH 6.5 @ 25°C

Heat Inactivation

65°C for 20 min

Methylation Sensitivity

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

Notes

  1. May exhibit star activity in NEBuffer 2.1 or CutSmart 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.

Faqs & Tech Tips

FAQs

  1. What is Star Activity and how can it be avoided?
  2. What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
  3. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
  4. How can I access the old NEBuffer Activity Chart?
  5. My restriction enzyme used to work well in the old NEBuffer but the new Performance chart indicates it has lower activity even though the only difference is the addition of BSA and removal of DTT to the new buffers. Why?
  6. Why is my Restriction Enzyme not cutting DNA?
  7. Why do I see a DNA smear on an agarose gel after a restriction digest?
  8. Why do I see additional DNA bands on my gel after a restriction digest?
  9. How many nucleotides do I have to add adjacent to the RE recognition site in order to get efficient cutting?
  10. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?

Protocols & Manuals

Protocols

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

Other Tools & Resources

Selection Charts

Usage Guildelines & Tips

Troubleshooting Guides

Restriction Enzyme Troubleshooting Guide

InteractiveTools

Citations

  • Tong C, Li H, Wang Y, Li X, Ou J, Wang D, Xu H, Ma C, Lang X, Liu G, Zhang B, Shi J. (2016) Construction of High-Density Linkage Maps of Populus deltoides × P. simonii Using Restriction-Site Associated DNA Sequencing. PLoS One. ; Mar 10;11(3), e0150692.
  • Mousavi M, Tong C, Liu F, Tao S, Wu J, Li H, Shi J. (2016) De novo SNP discovery and genetic linkage mapping in poplar using restriction site associated DNA and whole-genome sequencing technologies. BMC Genomics; Aug 18;17, 656.
  • Yang H, Wei CL, Liu HW, Wu JL, Li ZG, Zhang L, Jian JB, Li YY, Tai YL, Zhang J, Zhang ZZ, Jiang CJ, Xia T, Wan XC. (2016) Genetic Divergence between Camellia sinensis and Its Wild Relatives Revealed via Genome-Wide SNPs from RAD Sequencing. PLoS One. ; Mar 10;11(3), e0151424.
  • Fischer G, Azorsa F, Garcia FH, Mikheyev AS, Economo EP. (2015) Two new phragmotic ant species from Africa: morphology and next-generation sequencing solve a caste association problem in the genus Carebara Westwood. Zookeys.; Oct 5;(525), 77-105.
  • Bonatelli IA, Carstens BC, Moraes EM. (2015) Using Next Generation RAD Sequencing to Isolate Multispecies Microsatellites for Pilosocereus (Cactaceae). PLoS One. ; Nov 11;10(11), e0142602.
  • Xiao B, Tan Y, Long N, Chen X, Tong Z, Dong Y, Li Y. (2015) SNP-based genetic linkage map of tobacco (Nicotiana tabacum L.) using next-generation RAD sequencing. J Biol Res (Thessalon). ; Oct 6;22:11. ,
  • Zhang Q, Liu C, Liu Y, VanBuren R, Yao X, Zhong C, Huang H. (2015) High-density interspecific genetic maps of kiwifruit and the identification of sex-specific markers. DNA Res. ; Oct;22(5), 367-75.

Quality & Safety

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.

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 control's 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.