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Home Restriction Endonucleases Products MfeI

MfeI

This product will be discontinued on 12/15/2021 and is replaced with the high fidelity version, MfeI-HF™ (NEB #R3589).

NEB restriction endonuclease that recognizes the sequence C^AATT_G

Isoschizomers | Single Letter Code | Pronunciation:

100% activity in rCutSmart^™ Buffer (over 215 enzymes are available in the same buffer) allowing for easier double digests
High Fidelity (HF^®) version available (NEB #R3589) supplied with rCutSmart Buffer
Cut Site: C/AATTG

Catalog #	Concentration	Size
R0589S	10,000 units/ml	500 units
R0589L	10,000 units/ml	2,500 units
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Product Information

MfeI has a High Fidelity version, MfeI-HF® (NEB #R3589)

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 MfeI gene from Mycoplasma fermentans (N.F. Halden).

Reagents Supplied

The following reagents are supplied with this product:

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

R0589S -20

MfeI R0589SVIAL -20 1 x 0.05 ml 10,000 units/ml

rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X

R0589L -20

MfeI R0589LVIAL -20 1 x 0.25 ml 10,000 units/ml

rCutSmart™ Buffer B6004SVIAL -20 1 x 1.25 ml 10 X

Product Categories:: Restriction Endonucleases H M

Applications:: Restriction Enzyme Digestion

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 a total reaction volume of 50 µl.
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
NEBuffer™ r1.1: 75%
NEBuffer™ r2.1: 50%
NEBuffer™ r3.1: 10%
rCutSmart™ Buffer: 100%

Diluent Compatibility
- Diluent A
Storage Buffer

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

Heat Inactivation
No
Methylation Sensitivity

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

Isoschizomers

MfeI-HF
MunI
Related Products
Companion Products
Materials Sold Separately
- rCutSmart™ Buffer
Product Notes
1. MfeI is an isoschizomer of MunI.
2. Not sensitive to CpG, dcm, or dam methylation.
3. star activity may result from extended digestion.

Protocols, Manuals & Usage
- Protocols
  Optimizing Restriction Endonuclease Reactions
  
  Restriction Digest Protocol
  
  Double Digest Protocol with Standard Restriction Enzymes
- Usage Guidelines
  Activity at 37°C for Restriction Enzymes with Alternate Incubation Temperatures
  
  Activity of Restriction Enzymes in PCR Buffers
  
  Alteration of Apparent Recognition Specificities Using Methylases
  
  Cleavage Close to the End of DNA Fragments
  
  Digestion of Agarose-Embedded DNA: Info for Specific Enzymes
  
  Double Digests
  
  Heat Inactivation
  
  Restriction Endonucleases - Survival in a Reaction
  
  Restriction Enzyme Diluent Buffer Compatibility
  
  Restriction Enzyme Tips
  
  Single Letter Codes
  
  Star Activity
Tools & Resources
- Selection Charts
  Alphabetized List of Recognition Specificities
  
  Compatible Cohesive Ends and Generation of New Restriction Sites
  
  Dam-Dcm and CpG Methylation
  
  Frequencies of Restriction Sites
  
  Isoelectric Points (pI) for Restriction Enzymes
  
  Isoschizomers
  
  NEB Diluent and Buffer Table
  
  Recleavable Filled-in 5' Overhangs
  
  Time-Saver™ Qualified Enzymes
- Web Tools
  DNA Sequences and Maps Tool
  
  Double Digest Finder
  
  Enzyme Finder
  
  NEBcutter™ v2.0
  
  NEBioCalculator®
  
  REBASE®
FAQs & Troubleshooting
- FAQs
  Is MfeI inhibited by salt?
  
  Does MfeI have trouble cleavin PCR products?
  
  What is the molecular weight of MfeI?
  
  Does MfeI have single sites in common vectors?
  
  What is the activity of MfeI at 25°C?
  
  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?
  
  Why is my Restriction Enzyme not cutting DNA?
  
  Why do I see a DNA smear on an agarose gel after a restriction digest?
  
  Why do I see additional DNA bands on my gel after a restriction digest?
  
  How many nucleotides do I have to add adjacent to the RE recognition site in order to get efficient cutting?
  
  Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
  
  Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?
Citations & Technical Literature
- Citations
  
  Product Citation Tool
  
  Powered by Bioz See more details on Bioz
Quality, Safety & Legal
- Quality Control Assays
  
  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 & Change Notifications
  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]
  
  R0589S_L_v2
- 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]
  
  R0589S_L_v1_0081402
  
  R0589S_L_v1_0081407
  
  R0589S_L_v1_0081404
  
  R0589S_L_v1_0081409
  
  R0589S_L_v1_0081411
  
  R0589S_L_v1_0071212
  
  R0589S_L_v1_0081305
  
  R0589S_L v1_0081311
  
  R0589S_L_v1_0081503
  
  R0589S_L_v1_0081509
  
  R0589S_L_v1_0081510
  
  R0589S_L_v1_0081602
  
  R0589S_L_v1_0081604
  
  R0589S_L_v1_0081607
  
  R0589S_L_v2_0081611
  
  R0589S_L_v2_0081703
  
  R0589S_L_v2_0081705
  
  R0589S_L_v2_0081710
  
  R0589S_L_v2_0081712
  
  R0589S_L_v2_0081803
  
  R0589L_v2_10012877
  
  R0589S_v2_10012879
  
  R0589L_v2_10021707
  
  R0589L_v2_10028201
  
  R0589S_v2_10028199
  
  R0589L_v2_10034798
  
  R0589S_v2_10034800
  
  R0589L_v2_10043669
  
  R0589S_v2_10043671
  
  R0589L_v2_10052857
  
  R0589S_v2_10052860
  
  R0589L_v2_10064278
  
  R0589S_v2_10064280
  
  R0589L_v2_10073966
  
  R0589S_v2_10076924
  
  R0589L_v2_10079179
  
  R0589S_v2_10082142
  
  R0589L_v2_10088520
  
  R0589L_v2_10092124
  
  R0589S_v2_10088518
  
  R0589L_v2_10103873
  
  R0589S_v2_10104219
  
  R0589L_v2_10116126
  
  R0589S_v1_10116130
- Safety Data Sheets
  The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.
  
  MfeI
  
  rCutSmart™ Buffer
- Legal and Disclaimers
  
  This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).
  
  While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.
  
  For more information about commercial rights, please contact NEB's Global Business Development team at gbd@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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