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 sp. (strain 9°N™ FEN1 gene).
Reagents Supplied
The following reagents are supplied with this product:
|
Store at (°C) |
Concentration |
ThermoPol® Reaction Buffer |
-20 |
10X |
Description
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 sp. (strain 9°N™ FEN1 gene).
Reagents Supplied
The following reagents are supplied with this product:
|
Store at (°C) |
Concentration |
ThermoPol® Reaction Buffer |
-20 |
10X |
Advantages and Features
Applications
- Cleavage of flap DNA structure
Properties and 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 Temperature
-20°C
Storage Conditions
50 mM NaCl
10 mM Tris-HCl
10 mM MgCl2
1 mM DTT
pH 7.9 @ 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.
Related Products
Materials Sold Separately
References
1. Balakrishnon, L. et al. (2013). Annu. Rev. Biochem. 82, 119-138.
2. Greenough, L. et al. (2015). J. Biol. Chem. 20, 12514-12522.
3. Greenough, L. et al. (2015). Nucleic Acids Res. PubMedID: 26365239
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