Product Class: Other

T4 Phage β-glucosyltransferase (T4-BGT)
NEB4 cloned at NEB recombinant dil_B 37 65 Heat Epi

Product Introduction

T4 Phage β-glucosyltransferase specifically transfers the glucose moiety of uridine diphosphoglucose (UDP-Glc) to the 5-hydroxymethylcytosine (5-hmC) residues in double-stranded DNA, making beta-glucosyl-5-hydroxymethylcytosine (1,2).

  • Supplied with 10X NEBuffer 4, 50X UDP-Glucose (2 mM)
Catalog # Size Concentration
M0357S 500.0 units 10000 units/ml
M0357L 2500.0 units 10000 units/ml

Product Information

Description

Product Source

An E. coli strain that carries the cloned bgt gene from bacteriophage T4
This product is related to the following categories:
Epigenetic Analysis Products
This product can be used in the following applications:
5-hmC Detection & Analysis

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • M0357S     -20    
  • M0357L     -20    

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to protect 0.5 µg T4gt-DNA against cleavage by MfeI restriction endonuclease.

Reaction Conditions

1X NEBuffer™ 4
Supplement with 40 µM Uridine Diphosphate Glucose
Incubate at 37°C

1X NEBuffer™ 4
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
1 mM DTT
(pH 7.9 @ 25°C)

Diluent Compatibility

Storage Buffer

20 mM KPO4
200 mM NaCl
0.25 mM DTT
0.1 mM EDTA
50% Glycerol
pH 7 @ 25°C

Heat Inactivation

65°C for 10 minutes

Molecular Weight

Theoretical: 40666 kDa

Unit Assay Conditions

0.5 µg T4gt-DNA, 1X NEBuffer 4 and 40 µM UDP-Glucose in a 30 µl reaction. Incubate for 1 hour at 37°C followed by 10 minutes at 65°C. The extent of protection by T4 -BGT is determined by the addition of 20 µl 1X NEBuffer 4 and 10 units of MfeI. Incubation at 37°C for 30 minutes is followed by analysis on agarose gels.

Enzyme Properties
Activity in NEBuffers:
NEBuffer 1 100
NEBuffer 2 50
NEBuffer 3 50
NEBuffer 4 100% 

Survival in a Reaction: A minimum of 0.16 unit for 16 hours is required to protect 0.5 µg T4 gt-DNA against cleavage by MfeI.

References

  1. Josse, J. and Kornberg, A. (1962). J. Biol.Chem.. 237, 1968-1976.
  2. Tomaschewski, J. et al. (1985). Nucleic Acids Res. 13, 7551-7568.
  3. McNicol, L. A. et al. (1973). J. Mol. Biol. 15, 76,, 285-301.
  4. Szwagierczak, A. et al. (2010). Sensitive enzymatic quantification of 5-hydroxymethylcytosine in genomic DNA. Nucleic Acids Res. 38, e181.

Protocols, Manuals & Usage

Protocols

  1. Protocol for Glucosylation and digestion of Genomic DNA using AbaSI (#R0665)

Usage & Guidelines

FAQs & Troubleshooting

FAQs

  1. Will T4 Phage β-glucosyltransferase (T4-BGT) work in rCutSmart Buffer?

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 [email protected].

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.