Detection of 5-hydroxymethylcytosine by glycosylation
First Claim
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1. A method for detecting 5-hydroxymethylcytosine in a mammalian nucleic acid, the method comprising:
- contacting a sample comprising said mammalian nucleic acid with an enzyme that utilizes a labeled glucose or a labeled glucose-derivative donor substrate to add a labeled glucose molecule or a labeled glucose-derivative to a 5-hydroxymethylcytosine nucleic acid residue to generate a labeled glucosylated-5-hydroxymethylcytosine,wherein the enzyme is a beta-glucosyltransferase andwherein the labeled glucose or the labeled glucose-derivative donor substrate comprises a uridine diphosphate glucose (UDPG); and
treating the contacted sample with a TET enzyme or a catalytically active fragment thereof comprising SEQ ID NO;
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Abstract
Provided herein are methods and kits for detecting a modified cytosine.
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8 Claims
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1. A method for detecting 5-hydroxymethylcytosine in a mammalian nucleic acid, the method comprising:
contacting a sample comprising said mammalian nucleic acid with an enzyme that utilizes a labeled glucose or a labeled glucose-derivative donor substrate to add a labeled glucose molecule or a labeled glucose-derivative to a 5-hydroxymethylcytosine nucleic acid residue to generate a labeled glucosylated-5-hydroxymethylcytosine, wherein the enzyme is a beta-glucosyltransferase and wherein the labeled glucose or the labeled glucose-derivative donor substrate comprises a uridine diphosphate glucose (UDPG); and
treating the contacted sample with a TET enzyme or a catalytically active fragment thereof comprising SEQ ID NO;
1.- View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
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