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SacI

Catalog # Concentration Size List Price Quantity Your Price
R0156L 20000 units/ml 10000 units
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R0156M 100000 units/ml 10000 units
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R0156S 20000 units/ml 2000 units
Please Inquire
Catalog # Size List Price Your Price
R0156L 10000 units
R0156M 10000 units
R0156S 2000 units
Please Inquire
*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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    Restriction Enzyme Digest Protocol: Cutting Close to DNA End
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    Double Digestion with NEBcloner
SacI has a High Fidelity version SacI-HF® (NEB #R3156).

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.
Product Source
An E. coli strain that carries the SacI gene from Streptomyces achromogenes (ATCC 12767).

Properties & Usage

Unit Definition

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

Reaction Conditions

1X NEBuffer™ r1.1
Incubate at 37°C

1X NEBuffer™ r1.1
10 mM Bis-Tris-Propane-HCl
10 mM MgCl2
100 µg/ml Recombinant Albumin
(pH 7 @ 25°C)

Activity in NEBuffers

NEBufferâ„¢ r1.1: 100%
NEBufferâ„¢ r2.1: 50%
NEBufferâ„¢ r3.1: 10%
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
  • SacI is inhibited by salt concentrations > 10 mM. Mini-prep DNA containing residual salt is resistant to cleavage. A 70% alcohol wash or dialysis can be used to remove the salt.
  • 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.
  • 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.
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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