{"product_id":"neb-m0357s","title":"New England Biolabs, M0357S, T4 Phage β-glucosyltransferase (T4-BGT)","description":"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).  \u003cb\u003eRelated Categories\u003c\/b\u003e Hydroxymethylation Detection and Analysis,, Enzymatic Conversion for DNA Methylation Analysis,, DNA Methylation Analysis  \u003cb\u003eSpecification\u003c\/b\u003e \u003cb\u003eUnit Definition\u003c\/b\u003e One unit is defined as the amount of enzyme required to protect 0.5 µg T4gt-DNA against cleavage by MfeI restriction endonuclease.  \u003cb\u003eReaction Conditions\u003c\/b\u003e 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)  \u003cb\u003eDiluent Compatibility\u003c\/b\u003e Diluent B  \u003cb\u003eStorage Buffer\u003c\/b\u003e 20 mM KPO4 200 mM NaCl 0.25 mM DTT 0.1 mM EDTA 50% Glycerol pH 7 @ 25°C  \u003cb\u003eHeat Inactivation\u003c\/b\u003e 65°C for 10 minutes  \u003cb\u003eMolecular Weight\u003c\/b\u003e Theoretical: 40666 kDa  \u003cb\u003eUnit Assay Conditions\u003c\/b\u003e 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.  \u003cb\u003eFAQ\u003c\/b\u003e Q: Will T4 Phage β-glucosyltransferase (T4-BGT) work in rCutSmart Buffer? A: T4 Phage β-glucosyltransferase (T4-BGT) will work in rCutSmart Buffer. To see its % functional activity in rCutSmart, and that of other DNA Modifying enzymes in the cloning workflow, refer to the Activity of DNA Modifying Enzymes in rCutSmart® Buffer chart. ","brand":"New England Biolabs","offers":[{"title":"Default Title","offer_id":46835523485865,"sku":"M0357S","price":0.99,"currency_code":"USD","in_stock":true}],"url":"https:\/\/iright.com\/products\/neb-m0357s","provider":"Iright","version":"1.0","type":"link"}