COMPOSITION AND METHODS RELATED TO MODIFICATION OF 5-HYDROXYMETHYLCYTOSINE (5-hmC)

• US 20150056616A1
• Filed: 05/01/2014
• Published: 02/26/2015
• Est. Priority Date: 04/06/2010
• Status: Active Application

• Current Assignee: University of Chicago
• Original Assignee: University of Chicago
• Inventors: Song, Chunxiao, HE, Chuan
• Application Number:
  US14/267,727
• Publication Number:
  US 20150056616A1
• Time in Patent Office: Days
• US Class Current: 435/6.11
• CPC Class Codes: C12P 19/34 Polynucleotides, e.g. nucleic acids, oligoribonucleotides
• CPC Class Codes: C12Q 1/48 involving transferase
• CPC Class Codes: C12Q 1/6827 for detection of mutation or polymorphism
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 2521/531 Glycosylase
• CPC Class Codes: C12Q 2537/164 Methylation detection other then bisulfite or methylation sensitive restriction endonucleases
• CPC Class Codes: C12Q 2600/154 Methylation markers
• CPC Class Codes: C12Y 204/01028 Glucosyl-DNA beta-glucosyltransferase (2.4.1.28)

Abstract

The present invention relates generally to the field of molecular biology. More particularly, it concerns methods and compositions for detecting, evaluating, and/or mapping 5-hydroxymethyl-modified cytosine bases within a nucleic acid molecule.

Claims

1-162. -162. (canceled)

163. A kit comprising purified β

-glucosyltransferase and a modified uridine diphosphoglucose (UDP-Glu) molecule.

164. The kit of claim 163, further comprising one or more nucleic acid buffers.

165. The kit of claim 163, further comprising a polymerase.

166. The kit of claim 165, further comprising a composition comprising nucleotides for use with the polymerase.

167. The kit of claim 165, further comprising a restriction enzyme.

172. The kit of claim 163, wherein the modified UDP-Glu molecule comprises a linker, a detectable moiety, an isolation tag, a blocking moiety, or a functional moiety.

168-171. -171. (canceled)

173. A kit comprising purified β

-glucosyltransferase and a synthetically modified uridine diphosphoglucose (UDP-Glu) molecule, a polymerase, and one or more nucleic acid buffers.

174. The kit of claim 173, further comprising a composition comprising nucleotides for use with the polymerase.

175. The kit of claim 173, further comprising a restriction enzyme.

176. The kit of claim 173, wherein the synthetically modified UDP-Glu molecule comprises a linker, a detectable moiety, an isolation tag, a blocking moiety, or a functional moiety.

177. A kit comprising purified β

-glucosyltransferase and a synthetically modified uridine diphosphoglucose (UDP-Glu) molecule, a polymerase, and one or more nucleic acid buffers, wherein the synthetically modified UDP-Glu comprises a synthetic linker, a detectable moiety, an isolation tag, a blocking moiety, or a functional moiety.