IP Library Granted Patent US 8,771,939
Granted Patent B2
US 8,771,939 · App. 12/310,051 · Granted Jul 8, 2014

Method for methylation analysis of nucleic acid

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Quick Facts
Patent No.
US 8,771,939
App. No.
12/310,051
Granted
Jul 8, 2014
Kind
B2
Abstract

The present invention relates to a method for methylation analysis. It comprises the providing of a double stranded nucleic acid; its conversion, whereby unmethylated bases become distinguishable in their base-pairing behavior from methylated bases, and the analysis of both of the converted nucleic acid strands.

Claims (14)

1. A method for methylation analysis of a nucleic acid, comprising

providing a double stranded nucleic acid,

converting said nucleic acid in such a way that 5-methylcytosine remains unchanged, while unmethylated cytosine is converted to uracil or to another base that is distinguished by cytosine in its base-pairing behavior, said reaction leading to two different converted nucleic acid strands that are no longer complementary to each other,

analyzing both of the converted nucleic acid strands, wherein one of the two strands is analyzed in a methylation specific manner and the other of the two strands is analyzed in a non-methylation specific manner, wherein

(i) the methylation-specific manner comprises the analysis of the presence or absence of methylation of one or more cytosines; and

(ii) the non-methylation specific manner comprises the analysis of the copy number, deletion or amplification of one or more adjacent nucleotides in a corresponding, overlapping, adjacent or different section of the second strand or an analysis of SNP.

2. A method of claim 1 , wherein the converting of nucleic acid comprises a chemical reagent, bisulfite, an enzyme, or a cytidine-deaminase.

3. A method of claim 1 , wherein analyzing both of the converted nucleic acid strands comprises the analysis of corresponding, overlapping, adjacent or different sections of the strands of the originally provided nucleic acid.

4. A method of claim 1 , wherein analyzing both of the converted nucleic acid strands comprises

(a) the quantification of methylation or non-methylation of one or more CpG positions; and

(b) the quantification of converted nucleic acid; or the quantification of unconverted nucleic acids.

5. A method of claim 1 , wherein analyzing both of the converted nucleic acid strands comprises at least one method selected from the group consisting of amplification method, PCR method, isothermal amplification method, NASBA method, LCR method, methylation specific amplification method, MSP (Methylation Specific PCR) method, nested MSP method, HeavyMethyl™ method, methylation specific detection method, bisulfite sequencing method, detection by means of microarrays, detection by means of oligonucleotide microarrays, detection by means of restriction enzymes, simultaneous methylation specific amplification and detection method, real-time PCR, HeavyMethyl™ real time PCR method, MSP MethyLight™ method, MethyLightTM method, MethyLight™ A 1 go™ method, QM method, Headloop MethyLight™ method, HeavyMethyl™ MethyLight™ method, HeavyMethyl™ Scorpion™ method, MSP Scorpion™ method, Headloop Scorpion™ method, methylation sensitive primer extension, and Ms-SNuPE (Methylation-sensitive Single Nucleotide Primer Extension) method.

6. A method according to claim 1 , wherein genomic DNA is analyzed by real time PCR.

7. A method of claim 1 , wherein the methylation specific manner comprises a real time quantitative methylation method and wherein the non-methylation specific manner comprises a real time PCR method.

Assignments (2)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jun 7, 2024
From: EPIGENOMICS AG
To: NEW DAY DIAGNOSTICS LLC
Reel/Frame 067664/0713 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 9, 2009
From: TETZNER, REIMO; LEWIN, JOERN
To: EPIGENOMICS AG
Reel/Frame 022271/0952 →