Product Class: Restriction Endonuclease

AbaSI
neb31 cloned at NEB recombinant dil_C 25 65 Heat Epi

AbaSI was discontinued on September 15, 2023.

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

  • Catalog # R0665 was discontinued on September 15, 2023
NEB restriction endonuclease that recognizes 5-glucosylhydroxymethylcytosine (5ghmC) in double stranded DNA
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

  • Specificity to epigenetically-relevant DNA modifications (5-mC and 5-hmC)

  • Restriction Enzyme Cut Site: CNNNNNNNNNNN/NNNNNNNNNG

Bioz Badge Exists : True
Catalog # Size Concentration

Featured Videos

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Product Information

Description

Recognition sequence: x is ghmC, hmC, mC or C

AbaSI is a DNA modification-dependent endonuclease and it is an EpiMark® validated product. AbaSI recognizes 5-glucosylhydroxymethylcytosine (5ghmC) in double stranded DNA. It also recognizes 5-hydroxymethylcytocine (5hmC) but at a lower efficiency. It does not recognize DNA with 5-methylcytosin or un-modified cytosine. AbaSI selectively cleaves DNA that contains the modified bases, 5ghmC or 5hmC on one or both strands and introduces a double-stranded DNA break on the 3´ side away from the modified cytosine producing a 2-base or 3-base 3´-overhang. Sites with two 5ghmC on opposite strands are cleaved most efficiently; sites with one 5ghmC and another C or 5mC are cleaved less efficiently.

Product Source

An E. coli strain that carries the synthetic AbaSI gene from Acinetobacter baumannii SDF species.
This product is related to the following categories:

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 T4 wild-type phage DNA (fully ghmC-modified) in 1 hour at 25°C in a total reaction volume of 50 μl.

Reaction Conditions

1X rCutSmart™ Buffer
Supplement with 1 mM DTT
Incubate at 25°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: 25%
NEBuffer™ r2.1: 50%
NEBuffer™ r3.1: 50%
rCutSmart™ Buffer: 100%

Diluent Compatibility

Storage Buffer

100 mM KCl
10 mM Tris-HCl
1 mM DTT
0.1 mM EDTA
0.5% Tween® 20
0.5% IGEPAL® CA-630
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

Activity at Temperature

@37°C: 0%

Product Notes

  1. Optimization of the amount of enzyme for complete digestion of genomic DNA may be required.
  2. Not sensitive to CpG, dcm, or dam methylation.

Protocols, Manuals & Usage

Protocols

  1. Protocol for Glucosylation and digestion of Genomic DNA using AbaSI (#R0665)
  2. Restriction Digest Protocol
  3. Double Digest Protocol with Standard Restriction Enzymes

Usage & Guidelines

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. Does AbaSI cut hemi-glucosylated CpG site?
  2. How do I know my DNA is cut?
  3. How much enzyme should be used in digesting genomic DNA?
  4. Is this enzyme sensitive to dam, dcm or mammalian CpG methylation?
  5. 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.

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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