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AbaSI

Catalog # Concentration Size List Price Quantity Your Price
R0665S 10000 units/ml 1000 units
Please Inquire
Catalog # Size List Price Your Price
R0665S 1000 units
Please Inquire
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
  • 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

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    Double Digestion with NEBcloner
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.

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%


Companion Products
Notes
  • Optimization of the amount of enzyme for complete digestion of genomic DNA may be required.
  • Not sensitive to CpG, dcm, or dam methylation.
Citations
Additional Citations
  • Mooijman D, Dey S S, Boisset JC, Crosetto N, van Oudenaarden A (2016) Single-cell 5hmC sequencing reveals chromosome-wide cell-to-cell variability and enables lineage reconstruction Nat Biotechnol 34, 852-857.PubMedID: 27347753, DOI: 10.1038/nbt.3598
  • McKernan KJ, Spangler J, Zhang L, Tadigotla V, McLaughlin S, Warner J, Zare A, Boles RG (2014) Expanded genetic codes in next generation sequencing enable decontamination and mitochondrial enrichment PLoS One 9(5), e96492.PubMedID: 24788618, DOI: 10.1371/journal.pone.0096492
Publications
  • Sun Z, Terragni J, Borgaro JG, Liu Y, Yu L, Guan S, Wang H, Sun D, Cheng X, Zhu Z, Pradhan S, Zheng Y (2013). High-resolution enzymatic mapping of genomic 5-hydroxymethylcytosine in mouse embryonic stem cells Cell Rep. 3(2), 567-576.PubMedID: 23352666, DOI: 10.1016/j.celrep.2013.01.001
  • Wang H, Guan S, Quimby A, Cohen-Karni D, Pradhan S, Wilson G, Roberts RJ, Zhu Z, Zheng Y (2011). Comparative characterization of the PvuRts1I family of restriction enzymes and their application in mapping genomic 5-hydroxymethylcytosine Nucleic Acids Res. 39(21), 9294-305.PubMedID: 21813453, DOI: 10.1093/nar/gkr607
  • Horton JR, Borgaro JG, Griggs RM, Quimby A, Guan S, Zhang X, Wilson GG, Zheng Y, Zhu Z, Cheng X (2014). Structure of 5-hydroxymethylcytosine-specific restriction enzyme AbaSI in complex with DNA Nucleic Acids Res. 42(12), 7947-59.PubMedID: 24895434, DOI: 10.1093/nar/gku497
Quality Control Assay
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 Data Sheets
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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.

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