Product Class: Restriction Endonuclease

AleI-v2
neb31 cloned at NEB recombinant engineered dil_B 37 65 Heat CpG

This enzyme replaces AleI (NEB #R0634).

AleI-v2 has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10187675. Learn more.

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.

AleI-v2 is an engineered and improved version of AleI that recognizes the sequence CACNN^NNGTG
Isoschizomers | Single Letter Code

Product Introduction

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

  • Restriction Enzyme Cut Site: CACNN/NNGTG
Catalog # Size Concentration
R0685S 500.0 units 10000 units/ml
R0685L 2500.0 units 10000 units/ml

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 and modified AleI gene from Aureobacterium liquefaciens  (S.K. Degtyarev).
This product is related to the following categories:
Restriction Endonucleases: A Products
This product can be used in the following applications:
DNA Preparation,
Restriction Enzyme Digestion,
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: <10%
NEBuffer™ r2.1: <10%
NEBuffer™ r3.1: <10%
rCutSmart™ Buffer: 100%

Diluent Compatibility

Storage Buffer

10 mM Tris-HCl
200 mM NaCl
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: Impaired by Overlapping

Isoschizomers

OliI

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

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. 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. Which NEB restriction enzymes are supplied with Gel Loading Dye, Purple (6X)?
  6. Do degenerate recognition sites need to be palindromic?
  7. Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
  8. Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?

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

AleI-v2 has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10187675. 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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