EcoRI

Now comes supplied with 1 vial of Gel Loading Dye, Purple (6x). EcoRI has an HF version (NEB #R3101) supplied with CutSmart Buffer.
cloned at neb recombinant timesaver 5min unique buffer dil_C-grey incubation temp heat inactivation cpg blue white
EcoR-I-cutsite_1
Catalog #SizeConcentrationPriceQtyAdd to Cart
R0101S10,000 units20,000 units/ml$58.00Add to Cart
R0101T10,000 units100,000 units/ml$58.00Add to Cart
R0101L50,000 units20,000 units/ml$234.00Add to Cart
R0101M50,000 units100,000 units/ml$234.00Add to Cart
  
Categories:
Restriction Endonucleases: C-G,
Time-Saver™ Qualified Restriction Enzymes
Applications:
Restriction Enzyme Digestion

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-2010X

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

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

  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

  1. Optimizing Restriction Endonuclease Reactions
  2. Double Digest Protocol with Standard Restriction Enzymes
  3. Time-Saver Protocol for Restriction Enzyme Digests

Selection Charts

Usage Guidelines & Tips

Troubleshooting Guides

Interactive Tools

Citations

  • 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.
  • 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.
  • 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.
  • 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.
  • 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.

Quality Control

Quality Control Assays

The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page. Further information regarding NEB product quality can be found here.
  • Blue-White Screening (Terminal Integrity):
    A sample of DNA vector linearized with a 10-fold excess of a restriction endonuclease, religated and transformed into an E. coli strain expressing the LacZ beta fragment gene results in less than 1% white colonies.
  • Exonuclease Activity (Radioactivity Release):
    The product is tested in a reaction containing a radiolabeled mixture of single and double-stranded DNA. After incubation for 4 hours the exonuclease activity is determined by the % release of radioactive nucleotides.
  • Ligation and Recutting (Terminal Integrity):
    After an over-digestion of DNA with a restriction endonuclease the percentage of the DNA fragments ligated with T4 DNA ligase and the percentage that can be recut are determined by agarose gel electrophoresis.
  • Non-Specific DNase Activity (16 hour):
    The product is tested for non-specific nuclease degradation in a reaction containing a DNA substrate. After incubation for 16 hours there is no detectable degradation of the DNA substrate as determined by agarose gel electrophoresis.

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]

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]

Safety Data Sheet

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