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US 10,011,878 B2Granted
Compositions and methods for performing methylation detection assays
AssigneeExact Sciences Development Company, LLC
InventorsDavid Alan Ahlquist, William Russell Taylor, Douglas W. Mahoney, Graham P. Lidgard, Hatim T. Allawi, Abram Michael Vaccaro
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Abstract
Provided herein is technology relating to performing methylation assays. In particular, the technology relates to internal controls for methylation assays.
Claims8
A method of measuring the amount of methylated marker DNAs in a sample from a subject, the method comprising:
- a) extracting genomic DNA from a biological sample from a human subject;
- b) treating the extracted genomic DNA with bisulfite;
- c) measuring amounts of methylated marker DNA for two genes to seven genes selected from the group consisting of vimentin, septin 9, NDRG4, BMP3, VA V3, SFMBT2 897, and TFPI2a in the bisulfite-treated genomic DNA;
- d) measuring the amount of methylated ZDHHC1 DNA in the bisulfite-treated genomic DNA by amplifying a region of ZDHHC1 in the bisulfite treated genomic DNA using a pair of primers, wherein the amplifying produces an amplified product comprising a region of SEQ ID NO:
- 27, and contacting the amplified product with one or more oligonucleotides that comprise a sequence selected from the group consisting of SEQ ID NO:
- 28-32 and 34-36;
- e) calculating a value for the amount of each of said two to seven methylated marker genes measured in said bisulfite-treated genomic DNA as a percentage of the amount of methylated ZDHHC1 DNA measured in said bisulfite-treated genomic DNA, wherein the value indicates the amount of each of the two to seven methylated marker DNAs measured in the sample.
4 more elements
Dependencies are inferred from the claim text.