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NotI

Catalog # Concentration Size List Price Quantity Your Price
R0189L 10000 units/ml 2500 units $497.00
$447.30
R0189M 50000 units/ml 2500 units $497.00
$447.30
R0189S 10000 units/ml 500 units $120.00
$108.00
Catalog # Size List Price Your Price
R0189L 2500 units $497.00
$447.30
R0189M 2500 units $497.00
$447.30
R0189S 500 units $120.00
$108.00
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
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    Double Digestion with NEBcloner
NotI has a High Fidelity version NotI-HF® (NEB #R3189).

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 NotI gene from Nocardia otitidis-caviarum (ATCC 14630).
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  NotI R0189SVIAL -20 1 x 0.05 ml 10,000 units/ml
  NEBuffer™ r3.1 B6003SVIAL -20 1 x 1.25 ml 10 X
  Gel Loading Dye, Purple (6X) B7024AVIAL 25 1 x 0.5 ml 6 X
  NotI R0189LVIAL -20 1 x 0.25 ml 10,000 units/ml
  NEBuffer™ r3.1 B6003SVIAL -20 1 x 1.25 ml 10 X
  Gel Loading Dye, Purple (6X) B7024AVIAL 25 1 x 0.5 ml 6 X
  NotI R0189MVIAL -20 1 x 0.05 ml 50,000 units/ml
  NEBuffer™ r3.1 B6003SVIAL -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 pBC4 DNA in 1 hour at 37°C in a total reaction volume of 50 µl.

Reaction Conditions

1X NEBuffer™ r3.1
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: <10%>
NEBufferâ„¢ r2.1: 50%
NEBufferâ„¢ r3.1: 100%
rCutSmartâ„¢ Buffer: 25%

Diluent Compatibility

Storage Buffer

10 mM Tris-HCl
250 mM NaCl
1 mM DTT
0.1 mM EDTA
200 µg/ml Recombinant Albumin
50% Glycerol
0.15% Triton® X-100
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

Isoschizomers

CciNI
NotI-HF



Companion Products
Materials Sold Separately
Notes
  • Blocked by CpG methylation.
References
  • Samuelson, J.C., Morgan, R.D., Benner, J.S., Claus, T.E., Packard, S.L. and Xu, S.Y. (2006). Engineering a rare-cutting restriction enzyme: Genetic screening and selection of NotI variants. Nucleic Acids Research. 34, 796-805. PubMedID: 16456032
  • Zhang, P., Bao, Y., Higgins, L. and Xu, S.Y. (2007). Rational design of a chimeric endonuclease targeted to NotI recognition site. Protein Engineering, Design and Selection. 20, 497-504.
  • Lambert, A.R., Sussman, D., Shen, B., Maunus, R., Nix, J., Samuelson, J., Xu, S.Y. and Stoddard, B.L. (2008). Structures of the Rare-Cutting Restriction Endonuclease NotI Reveal a Unique Metal Binding Fold Involved in DNA Binding. Structure. 16, 558-569.
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.
Change Notifications

NotI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10232248. 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 Data Sheets
Legal And Disclaimer

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 busdev@neb.com.

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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