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NsiI-HF®

Catalog # Concentration Size List Price Quantity Your Price
R3127L 20000 units/ml 5000 units $445.00
$400.50
R3127S 20000 units/ml 1000 units $108.00
$97.20
Catalog # Size List Price Your Price
R3127L 5000 units $445.00
$400.50
R3127S 1000 units $108.00
$97.20
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

High-Fidelity (HF®) restriction enzymes have the same specificity as native enzymes, but have been engineered for significantly reduced star activity and performance in a single buffer (rCutSmart™ Buffer).  All HF-restriction enzymes come with Gel Loading Dye, Purple (6X).  Enjoy the enhanced performance and added value of our engineered enzymes at the same price as the native enzyme:

  • Engineered for improved performance
  • 100% activity in rCutSmart Buffer
  • Time-Saver™ qualified for digestion in 5-15 minutes
  • Reduced star activity
  • Supplied with 1 vial of Gel Loading Dye, Purple (6X)
  • Restriction Enzyme Cut Site: ATGCA/T
Featured Videos
View Video Library
  • Monarch_MiniprepProtocol_VideoThumb
    Reduce Star Activity with High-Fidelity Restriction Enzymes
  • Monarch_MiniprepProtocol_VideoThumb
    Standard Protocol for Restriction Enzyme Digests
  • Monarch_MiniprepProtocol_VideoThumb
    NEB® TV Ep. 15 – Applications of Restriction Enzymes
  • Monarch_MiniprepProtocol_VideoThumb
    Restriction Enzyme Digest Protocol: Cutting Close to DNA End
  • Monarch_MiniprepProtocol_VideoThumb
    Restriction Enzyme Digestion Problem: DNA Smear on Agarose Gel
  • Monarch_MiniprepProtocol_VideoThumb
    Why is My Restriction Enzyme Not Cutting DNA?
  • Monarch_MiniprepProtocol_VideoThumb
    Restriction Enzyme Digest Problem: Too Many DNA Bands
  • Monarch_MiniprepProtocol_VideoThumb
    Double Digestion with NEBcloner

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 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.


NEB extensively performs quality controls on all standard and high-fidelity (HF) restriction enzymes. Examples of nuclease contamination studies for some of our HF restriction enzymes are shown below.

Restriction Enzyme Competitor Study: Nuclease Contamination


EcoRI, NotI, and BamHI from multiple suppliers were tested in reactions containing a fluorescent labeled single stranded, double stranded blunt, 3’overhang or 5’ overhang containing oligonucleotides. The percent degradation is determined by capillary electrophoresis and peak analysis. The resolution is at the single nucleotide level.
Product Source
An E. coli strain that carries the clone and modified NsiI gene from Neisseria sicca (ATCC 29256).
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  NsiI-HF® R3127SVIAL -20 1 x 0.05 ml 20,000 units/ml
  rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X
  Gel Loading Dye, Purple (6X) B7024AVIAL 25 1 x 0.5 ml 6 X
  NsiI-HF® R3127LVIAL -20 1 x 0.25 ml 20,000 units/ml
  rCutSmart™ Buffer B6004SVIAL -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 λ DNA in 1 hour at 37°C in 50 µl of reaction buffer.

Reaction Conditions

1X rCutSmart™ Buffer
Incubate at 37°C

1X rCutSmart™ Buffer
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
100 µg/ml Recombinant Albumin
(pH 7.9 @ 25°C)

Activity in NEBuffers

NEBufferâ„¢ r1.1: <10%>
NEBufferâ„¢ r2.1: 20%
NEBufferâ„¢ r3.1: <10%>
rCutSmartâ„¢ Buffer: 100%

Diluent Compatibility

Storage Buffer

10 mM Tris-HCl
300 mM NaCl
1 mM DTT
0.1 mM EDTA
500 µg/ml Recombinant Albumin
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

80°C for 20 minutes

Methylation Sensitivity

dam methylation: Not Sensitive
dcm methylation: Not Sensitive
CpG Methylation: Not Sensitive



Companion Products
Materials Sold Separately
Notes
  • Not sensitive to CpG, dcm, or dam methylation.
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

NsiI-HF has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10244729. All subsequent (higher number) lots will contain rAlbumin. For additional information, please visit 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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