Product Class: Restriction Endonuclease

icon_cloned icon_recomb engineered icon_CutSmart dil_B-grey icon_incTemp37 icon_HI_65 icon_cpg
Item# Description List Price Web Price Qty
R0685L AleI-v2 - 2,500 units $388.00
R0685S AleI-v2 - 500 units $95.00
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at

This enzyme replaces AleI (NEB #R0634).

    Product Introduction

    Having supplied restriction enzymes to the research community for over 40 years, NEB has earned the reputation of being the leader in enzyme technologies. Working continuously to be worth of that distinction, NEB strives to develop enzyme of the highest purity and unparalleled quality.

    100% activity in CutSmart® Buffer (over 210 enzymes are available in the same buffer) allowing for easier double digests

    Product Information


    AleI-v2 is an engineered and improved version of AleI. (NEB uses the designation "v2" or "HF" in the name to indicate engineered enzymes.)

    Product Source

    An E. coli strain that carries the cloned AleI gene from Aureobacterium liquefaciens  (S.K. Degtyarev).

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C) Concentration
    CutSmart® Buffer -20 10X
    Product Categories:
    Restriction Endonucleases Products
    DNA Preparation,
    Restriction Enzyme Digestion,
    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 CutSmart® Buffer
    Incubate at 37°C

    1X CutSmart® Buffer
    50 mM Potassium Acetate
    20 mM Tris-acetate
    10 mM Magnesium Acetate
    100 µg/ml BSA
    (pH 7.9 @ 25°C)

    Activity in NEBuffers

    CutSmart® Buffer: 100%
    NEBuffer™ 1.1: 10%
    NEBuffer™ 2.1: 10%
    NEBuffer™ 3.1: 10%

    Diluent Compatibility

    Storage Temperature


    Storage Conditions

    10 mM Tris-HCl
    200 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 min

    Methylation Sensitivity

    dam methylation: Not Sensitive
    dcm methylation: Not Sensitive
    CpG Methylation: Impaired by Overlapping

    Product Notes

    1. AleI-v2 is an isoschizomer of OliI.
    2. This enzyme is impaired by Cpg methylation. More information can be found at "Dam-Dcm and CpG Methylation."
    3. AleI-v2 has the same specificity as AleI (NEB #R0634), but it has been engineered for increased stability at -20C.

    Protocols, Manuals & Usage


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

    Usage & Guidelines

    • Activity of Restriction Enzymes in PCR Buffers
    • Cleavage Close to the End of DNA Fragments
    • Effects of CpG Methylation on Restriction Enzyme Cleavage
    • Heat Inactivation
    • NEBuffer Activity/Performance Chart with Restriction Enzymes
    • Restriction Endonucleases - Survival in a Reaction
    • Restriction Enzyme Diluent Buffer Compatibility

    Tools & Resources

    Selection Charts

    • Alphabetized List of Recognition Specificities
    • Dam-Dcm and CpG Methylation
    • Frequencies of Restriction Sites
    • Isoschizomers
    • Why Choose Recombinant Enzymes?

    Web Tools

    Faqs & Troubleshooting


    1. What is the difference between AleI (NEB #R0634) and AleI-v2 (NEB #R0685)?
    2. When is star activity a concern?
    3. 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?
    4. Is Gel Loading Dye, Purple (6X) or Gel Loading Dye, Purple (6X), no SDS compatible with other DNA binding dyes such as SYBR® and GelRed™ during gel electrophoresis?
    5. Why is my Restriction Enzyme not cutting DNA?
    6. Why do I see additional DNA bands on my gel after a restriction digest?
    7. Why do I see a DNA smear on an agarose gel after a restriction digest?
    8. How many nucleotides do I have to add adjacent to the RE recognition site in order to get efficient cutting?
    9. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?
    10. Do degenerate recognition sites need to be palindromic?


    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.


    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

    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 [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.