Product Class: Restriction Endonuclease

MlyI
Gray property icon for rCutSmart Gray property icon for cloned at NEB recombinant time saver Gray property icon for dilA Gray property icon for incubation temperature 37° Gray property icon for heat inactivation 65°

MlyI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10156747. Learn more.

NEB restriction endonuclease that recognizes the sequence GAGTCNNNNN_^
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
  • Restriction Enzyme Cut Site: GAGTC(5/5)

Bioz Badge Exists : True
Catalog # Size Concentration
R0610S 1000 units 10000 units/ml
R0610L 5000 units 10000 units/ml

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Product Information

Description

Product Source

An E. coli strain that carries the MlyI gene from Micrococcus lylae (NBL 2048).
This product is related to the following categories:
Restriction Endonucleases H M,
Time-Saver Qualified Restriction Enzymes,
This product can be used in the following applications:
Fast Cloning: Accelerate your cloning workflows with reagents from NEB,
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 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: 50%
NEBuffer™ r2.1: 50%
NEBuffer™ r3.1: 10%
rCutSmart™ Buffer: 100%

Diluent Compatibility

Storage Buffer

10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
200 µg/ml Recombinant Albumin
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

Product Notes

  1. MlyI is an isoschizomer of PleI that generates blunt-ended DNA fragments. Nt.BstNBI also recognizes the sequence GAGTC but only cleaves one strand of the DNA substrate.
  2. 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.
  3. Not sensitive to CpG, dcm, or dam methylation.

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. Is extended digestion of MlyI recommended?
  2. How does MlyI differ from its neoschizomers, PleI and Nt.BstNBI?
  3. Do I have to set-up digests with Time-Saver™ qualified enzymes for 5-15 minutes? Can I digest longer?
  4. 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?
  5. Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
  6. 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

Effective March 24, 2026, exonuclease specification updated.

MlyI has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10156747. All subsequent (higher number) lots will contain rAlbumin. 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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