Product Class: Restriction Endonuclease

SfiI
neb31 cloned at NEB recombinant time saver 2site dil_C 50 No CpG dcm

We are excited to announce that all reaction buffers are now BSA-free. NEB began switching our BSA-containing reaction buffers in April 2021 to buffers containing Recombinant Albumin (rAlbumin) for restriction enzymes and some DNA modifying enzymes. Find more details at www.neb.com/BSA-free.

NEB restriction endonuclease that recognizes the sequence GGCCN_NNN^NGGCC

Product Introduction

  • Time-Saver™ qualified for digestion in 5-15 minutes
  • 100% activity in rCutSmartBuffer (over 210 enzymes are available in the same buffer) simplifying double digests
  • Supplied with 1 vial of Gel Loading Dye, Purple (6X)
  • 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.

  • Cut Site: GGCCNNNN/NGGCC

Catalog # Size Concentration
R0123S 3000.0 units 20000 units/ml
R0123L 15000.0 units 20000 units/ml

Product Information

Description

SfiI crystals (Lydia Dorner and Ira Schildkraut, New England Biolabs)

SfiI crystals (Lydia Dorner and Ira Schildkraut, New England Biolabs)

Product Source

An E. coli strain that carries the SfiI gene from Streptomyces fimbriatus (ATCC 15051).
This product is related to the following categories:
Restriction Endonucleases S Products,
Time-Saver Qualified Restriction Enzymes Products,
This product can be used in the following applications:
Restriction Enzyme Digestion

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

Properties & Usage

Unit Definition

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

Reaction Conditions

1X rCutSmart™ Buffer
Incubate at 50°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

NEBuffer™ r1.1: 25%
NEBuffer™ r2.1: 100%
NEBuffer™ r3.1: 50%
rCutSmart™ Buffer: 100%

Diluent Compatibility

Storage Buffer

10 mM Tris-HCl
250 mM NaCl
0.1 mM EDTA
200 µg/ml BSA
50% Glycerol
0.15% Triton® X-100
1 mM DTT
pH 7.4 @ 25°C

Heat Inactivation

No

Methylation Sensitivity

dam methylation: Not Sensitive
dcm methylation: Impaired by Overlapping
CpG Methylation: Blocked by Some Combinations of Overlapping

Activity at Temperature

@37°C: 10%

Product Notes

  1. SfiI requires two copies of its recognition sequence for cleavage to occur. The two sites can be on either the same or different DNA molecules. Wertzell L.M. et al., (1995) J. Mol. Biol. 248:581-595.

    For more information and recommendations on working with enzymes requiring multi-sites, please visit Restriction Enzyme Cleavage: ‘single-site’ enzymes and ‘multi-site’ enzymes
  2. For enzymes that cannot be heat-inactivated, we recommend using a column for cleanup (such as the Monarch® PCR & DNA Cleanup Kit), or running the reaction on an agarose gel and then extracting the DNA (we recommend Monarch Gel Extraction Kit), or performing a phenol/chloroform extraction.
  3. 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.
  4. Impaired by overlapping dcm methylation or blocked by some combinations and overlapping CpG methylation.

Protocols, Manuals & Usage

Protocols

  1. Optimizing Restriction Endonuclease Reactions
  2. Restriction Digest Protocol
  3. Double Digest Protocol with Standard Restriction Enzymes

Usage & Guidelines

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. What is the minimum distance needed between two SfiI sites? Can they follow each other directly?
  2. Can I achieve digestion of a plasmid with a single SfiI site?
  3. Do degenerate recognition sites need to be palindromic?
  4. How can this enzyme be inactivated?
  5. Why isn't the enzyme cutting completely?
  6. Can I add an oligo to my reaction so that two sites are available for efficient cutting?
  7. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
  8. I tested your restriction enzyme on the substrate DNA recommended by NEB, and it appears to be active, however it does not digest my DNA. What could be the reason?
  9. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?
  10. Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
  11. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?

Troubleshooting

Quality, Safety & Legal

Quality Assurance Statement

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 DataSheets

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

Legal and Disclaimers

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 [email protected].

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.