Introduction
T4 β-glucosyltransferase (T4 BGT) is a DNA-modifying enzyme encoded by bacteriophage T4 that catalyzes the transfer of glucose (Glc) from uridine diphosphoglucose (UDP-Glc) to 5-hydroxymethylcytosine (5-hmC) residues in double-stranded DNA, resulting in the formation of β-glucosyl-5- hydroxymethylcytosine.
The enzyme is available in 500 and 2,500 unit sizes at a concentration of 10 U/µL. The enzyme is supplied with a 10× Reaction Buffer.
Applications
- Glucosylation or immunodetection of 5-hmC DNA.
- Differentiation of 5-hmC from 5-mC.
- 5-hmC containing DNA enrichment.
- Labeling of 5-hmC residues using a radioactive UDP-glucose donor.
Product Specifications
- Storage Buffer: 50 mM Tris-HCl (pH 7.5 at 25 oC), 100 mM NaCl, 0.1 mM EDTA, 1 mM DTT, 0.1% Triton X-100, and 50% (v/v) glycerol.
- 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.
- Protection Unit Assay Conditions: 0.5 µg T4gt-DNA, 1X reaction buffer 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 β-glucosyltransferase is determined by the addition of 20 µL 1X reaction buffer and 10 units of MfeI. Incubation at 37°C for 30 minutes is followed by analysis on agarose gels.
To Order
Related Products
Related Products
Related Products
- Tn5 Transposase
- T4 β-Glucosyltransferase
- T4 DNA Ligase
- Quick T4 DNA Ligation Kit
- Blunt/TA Ligase Master Mix
- Terminal Transferase (TdT)
- Benzonase Nuclease