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EcoP15I

Catalog # Concentration Size List Price Quantity Your Price
R0646S 10000 units/ml 500 units $112.00
$100.80
Catalog # Size List Price Your Price
R0646S 500 units $112.00
$100.80
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
  • This product is supplied with one vial of 10X ATP solution (10mM concentration, NEB #B6101S, not sold separately)
  • Time-Saver™ qualified for digestion in 5-15 minutes

  • Requires two or more sites for cleavage. Please review the enzyme list and recommendations in the following table as well as the background information article.

  • Restriction Enzyme Cut Site: CAGCAG(25/27)

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Product Source
An E. coli strain that carries the cloned EcoP15 l res-mod genes from plasmid pSHl180 (D.N. Rao)
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  EcoP15I R0646SVIAL -20 1 x 0.05 ml 10,000 units/ml
  NEBuffer™ r3.1 B6003SVIAL -20 1 x 1.25 ml 10 X
  10X ATP B6101SVIAL -20 1 x 0.3 ml 10 X

Properties & Usage

Unit Definition

One unit is defined as the amount enzyme required to digest 1 µg of pUC19 DNA in 1 hour at 37°C in a total reaction volume of 50 µl.

Reaction Conditions

1X NEBuffer™ r3.1
Supplement with 1X ATP
Incubate at 37°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

NEBufferâ„¢ r1.1: 75%
NEBufferâ„¢ r2.1: 100%
NEBufferâ„¢ r3.1: 100%
rCutSmartâ„¢ Buffer: 100%

Diluent Compatibility

Storage Buffer

10 mM Tris-HCl
100 mM NaCl
1 mM DTT
0.1 mM EDTA
200 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: Not Sensitive



Companion Products
Notes
  • ATP is required for DNA cleavage. Efficient cleavage requires the presence of two inversely oriented recognition sites. A head to head orientation is preferred. The "head" of the sequence is defined as the dG at the 3´ end of the CAGCAG strand (top strand). Cleavage efficiency is also affected by the distance between the two sites.
  • EcoP15I requires two copies of its recognition sequence for cleavage to occur, with limited cleavage on plasmids with a single site.
  • 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.
  • Requires two or more sites for cleavage. Please review the enzyme list and recommendations in the following table as well as the background information article.
  • Not sensitive to CpG, dcm, or dam methylation.
References
  • Elisabeth Möncke-Buchner, Maja Rothenberg, Stefanie Reich, Katja Wagenführ, Hideo Matsumura, Ryohei Terauchi, Detlev H. Krüger, and Monika Reuter (2009). Functional Characterization and Modulation of the DNA Cleavage Efficiency of Type III Restriction Endonuclease EcoP15I in Its Interaction with Two Sites in the DNA Target. J. Mol. Biol. 387, 1309–1319.
Citations
Additional Citations
  • Bhattacharyya, S., Yu, Y., Suzuki, M., Campbell, N., Mazdo, J., Vasanthakumar, A., et al. (2013) Genome-wide hydroxymethylation tested using the HELP-GT assay shows redistribution in cancer Nucleic Acids Res DOI: doi:10.1093/nar/gkt601
Quality Control Assay
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 controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]
Safety Data Sheets
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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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