Product Class: Other

5-hydroxymethyluridine DNA Kinase
NEBU cloned at NEB recombinant 37 80 Heat

Product Introduction

  • 5-hydroxymethyluridine DNA Kinase (5-HMUDK) transfers the gamma phosphate from ATP to the hydroxymethyl moiety of 5-hydroxymethyluridine in polymeric DNA
  • This is an Enzyme for Innovation (EFI). 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.
Catalog # Size Concentration
M0659S 1000.0 units 20000 units/ml

Product Information

Description

5-hydroxymethyluridine DNA Kinase (5-HMUDK) transfers the gamma phosphate from ATP to the hydroxymethyl moiety of 5-hydroxymethyluridine in polymeric DNA.

Specificity

Product Source

The 5-hydroxymethyluridine DNA Kinase enzyme is isolated from a strain of E. coli which contains the htdk gene from Pseudomonas aeruginosa bacteriophage M6.
This product is related to the following categories:
Kinases Products
This product can be used in the following applications:
DNA End Modification,
Phosphorylation (Kinase)

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 protect 1 µg of Bacillus subtilis bacteriophage SP8 genomic DNA in 30 minutes at 37°C in a total reaction volume of 20 µl against subsequent cleavage by NcoI-HF restriction endonuclease.

Reaction Conditions

1X T4 DNA Ligase Reaction Buffer
Incubate at 37°C

1X T4 DNA Ligase Reaction Buffer
50 mM Tris-HCl
10 mM MgCl2
1 mM ATP
10 mM DTT
(pH 7.5 @ 25°C)

Storage Buffer

300 mM NaCl
10 mM Tris-HCl
0.1 mM EDTA
1 mM DTT
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

80°C for 10 minutes

Unit Assay Conditions

1X T4 DNA Ligase Reaction Buffer, 1 µg of Bacillus subtilis bacteriophage SP8 genomic DNA in a total reaction volume of 20 µl. The reaction mix is incubated for 30 minutes at 37°C and then incubated at 80°C for 10 minutes. Confirmation of phosphorylation is determined by protection of subsequent cleavage by NcoI-HF restriction endonuclease. The reaction mix is treated with 2 µl of 10X rCutSmart Buffer and 20 units of NcoI-HF restriction endonuclease. Incubation at 37°C for 30 minutes is followed by analysis on an agarose gel.    

References

  1. Weigele, P.R., et al. (2017). FASEB. 31,

Protocols, Manuals & Usage

Protocols

  1. Protocol for a substrate with 5-hydroxymethyluridine such as bacteriophage SP8 genomic DNA (M0659)

FAQs & Troubleshooting

FAQs

  1. What DNA substrates has 5-hydroxymethyluridine DNA Kinase (5-HMUDK) been tested on?
  2. What is the activity of 5-hydroxymethyluridine DNA Kinase (5-HMUDK) at other temperatures, buffers, etc?
  3. Can I purchase large amounts of an existing Enzyme for Innovation?
  4. What are Enzymes for Innovation?

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.

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.