This enzyme replaces AleI (NEB #R0634).
    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
    $349.20
    ADD TO CART
    R0685S AleI-v2 - 500 units $95.00
    $85.50
    ADD TO CART
    *On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
    Ale-I-cutsite_1_v1_000007

    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

    Description

    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 10 X
    Product Categories:
    Restriction Endonucleases Products
    Applications:
    DNA Preparation,
    Restriction Enzyme Digestion,
    Restriction Enzyme Digestion

    Properties and 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

    -20°C

    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

    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.

    Faqs & Tech Tips

    FAQs

    1. What is the difference between AleI (NEB #R0634) and AleI-v2 (NEB #R0685)?
    2. When is star activity a concern?
    3. What effect does BSA have on the performance of NEB’s restriction enzymes when included in the new buffers?
    4. How can I access the old NEBuffer Activity Chart?
    5. 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?
    6. 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?
    7. Why is my Restriction Enzyme not cutting DNA?
    8. Why do I see additional DNA bands on my gel after a restriction digest?
    9. Why do I see a DNA smear on an agarose gel after a restriction digest?
    10. How many nucleotides do I have to add adjacent to the RE recognition site in order to get efficient cutting?
    11. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?
    12. Do degenerate recognition sites need to be palindromic?

    Protocols & Manuals

    Protocols

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

    Other Tools & Resources

    Selection Charts

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

    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

    Interactive Tools

    Quality & Safety

    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.

    AleI-v2
    CutSmart® Buffer