Product Class: Restriction Endonuclease

MmeI
neb31 cloned at NEB recombinant time saver 2site dil_B 37 65 Heat CpG

MmeI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10263881. Learn more.

SAM now included in enzyme formulation – no longer supplied as a separate vial.

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 TCCRACNNNNNNNNNNNNNNNNNN_NN^
Isoschizomers | Single Letter Code

Product Introduction

  • Time-Saver™ qualified for digestion in 5-15 minutes
  • 100% activity in rCutSmart™ Buffer (over 210 enzymes are available in the same buffer) allowing for easier double digests
  • Type IIS restriction enzymes recognize asymmetric DNA sequences and cleave outside of their recognition sequence
  • 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.
  • Restriction Enzyme Cut Site: TCCRAC(20/18)
Catalog # Size Concentration
R0637S 100.0 units 2000 units/ml
R0637L 500.0 units 2000 units/ml

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Product Information

Description

Product Source

An E. coli strain that carries the MmeI gene from Methylophilus methylotrophus.
This product is related to the following categories:
Restriction Endonucleases H M 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 MmeI required to digest 1 µg of ΦX174 RF I 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

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

Diluent Compatibility

Storage Buffer

10 mM Tris-HCl
300 mM NaCl
0.1 mM EDTA
500 µg/ml Recombinant Albumin
50% Glycerol
0.32 mM S-adenosylmethionine (SAM)
1 mM DTT
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 by Overlapping

Product Notes

  1. Excess MmeI blocks cleavage.
  2. Reactions using MmeI should be done at or near stoichiometric concentrations.
  3. Complete cleavage occurs within 15 minutes at 37°C.
  4. Significant cleavage occurs on ice and at 50°C.
  5. MmeI activity is inhibited by high ionic strength (> 200 mM).
  6. MmeI prefers two copies of its recognition sequence for cleavage to occur, with limited cleavage on plasmids with a single site.
  7. Based on the stability of the enzyme in the reaction, incubations longer than 1 hr will not result in improved digestion, unless additional enzyme is added. Please refer to Restriction endonuclease survival in a reaction for more information regarding this topic.
  8. 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.
  9. Blocked by overlapping CpG methylation.
  10. MmeI requires S-adenosylmethionine (SAM) in reaction mixture for optimal activity (SAM is supplied in the enzyme formulation).

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. Is there any variability in the distance of cutting from the MmeI cut site which is indicated as 20/18?
  2. I am observing partial digestion when using MmeI. What could be the reason?
  3. Is MmeI sensitive to CpG methylation?
  4. Why can a cleaved MmeI site religate but not be re-cut?
  5. Is MmeI cutting exact or is the spacing variable?
  6. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
  7. 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?
  8. Will incubation overnight improve my digest?
  9.  Does MmeI require SAM for activity?
  10. 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.

Specification Change Notifications

MmeI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10263881. 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 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 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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