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Thermostable FEN1

Catalog # Concentration Size List Price Quantity Your Price
M0645S 32000 units/ml 1600 units $123.00
$110.70
Catalog # Size List Price Your Price
M0645S 1600 units $123.00
$110.70
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

  • Thermostable DNA and RNA endonuclease
  • Catalyzes the cleavage of 5' DNA and RNA flaps from branched double stranded DNA substrates, creating a 5' phosphate terminus
  • FEN 1 products can be ligated by DNA ligase to create double stranded DNA
  • Heat inactivation not possible
  • This is an Enzyme for Innovation. EFI is a project initiated by NEB to provide unique enzymes to the scientific community in the hopes of enabling the discovery of new and innovative applications. These enzymes have interesting properties and unique specificities
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    What are endonucleases and their applications?
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Thermostable Flap Endonuclease 1, FEN1, catalyzes the cleavage of 5´ DNA flaps from branched double stranded DNA substrates, creating a 5´ phosphate terminus. FEN1 products can be ligated by DNA ligase to create double stranded DNA. In vivo, FEN1 is an essential component of the Okazaki fragment maturation pathway, and also plays a role in base excision repair.

Reaction Schematic Thermostable FEN1 recognizes and cleaves 5´ DNA flaps generating a ligatable 5´ phosphate terminus.
Product Source
An E. coli strain, which carries the cloned Thermococcus 9°N™ FEN1 gene.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  Thermostable FEN1 M0645SVIAL -20 1 x 0.05 ml 32,000 units/ml
  ThermoPol® Reaction Buffer B9004SVIAL -20 1 x 1.5 ml 10 X
Application Features
  • Cleavage of flap DNA structure

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to cleave 10 pmol of 5´ flap containing oligonucleotide substrate in a total reaction volume of 10 µl for 10 min at 65°C.

Reaction Conditions

1X ThermoPol® Reaction Buffer
Incubate at 65°C

1X ThermoPol® Reaction Buffer
20 mM Tris-HCl
10 mM (NH4)2SO4
10 mM KCl
2 mM MgSO4
0.1% Triton® X-100
(pH 8.8 @ 25°C)

Storage Buffer

10 mM Tris-HCl
1 mM DTT
100 mM KCl
0.1 mM EDTA
0.1% Triton® X-100
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

No

Unit Assay Conditions

1X ThermoPol Reaction Buffer containing 0.2 pmol of fluorescently labeled oligonucleotide in a total reaction volume of 10 µl.

Materials Sold Separately
References
  • Balakrishnon, L. et al. (2013). Annu. Rev. Biochem. 82, 119-138.
  • Greenough, L. et al. (2015). J. Biol. Chem. 20, 12514-12522.
  • Greenough, L. et al. (2015). Nucleic Acids Res. DOI: doi:10.1093/nar/gkv899 PubMedID: 26365239
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
Legal And Disclaimer

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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