Unlock instant, AI-driven research and patent intelligence for your innovation.

Base specific cleavage of methylation-specific amplification products in combination with mass analysis

A technology of amplification products and methylation, which is used in the determination/inspection of microorganisms, biochemical equipment and methods, fermentation, etc., and can solve the problems of labor consumption and a large amount of high-quality genomic DNA.

Inactive Publication Date: 2008-07-30
SEQUENOM INC
View PDF37 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Some methods are prone to false positive results, thus limiting methylation assessment to a single cytosine site per analysis
Other methods require large amounts of high-quality genomic DNA, are labor-intensive, and are prone to false-positive results

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Base specific cleavage of methylation-specific amplification products in combination with mass analysis
  • Base specific cleavage of methylation-specific amplification products in combination with mass analysis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment I

[0650] The following example illustrates a method for identifying the molecular methylation status of a target nucleic acid molecule.

[0651] method

[0652] bisulfite treatment

[0653] Bisulfite treatment was performed according to the method given in Paulin et al. Nucl. Acids Res. 26:5009-5011 (1998). Genomic DNA (2 μg) was digested with restriction endonucleases, then denatured by adding 3M sodium hydroxide and incubated at 37°C for 15 min. Add 6.24M urea / 2M meta-bisulfite (4M bisulfite) solution and 10mM sodium hydroquinone solution to the denatured DNA to achieve the final concentration: 5.36M urea, 3.44M bisulfite, 0.5 mM hydroquinone. The DNA solution was placed in a 0.5 ml tube and covered with mineral oil, and 20 cycles were performed in a PCR machine, with the tube repeatedly heated to 95°C for 30 seconds and incubated at 55°C for 15 min. In addition to desalting the DNA using a Bresaclean DNA purification procedure (Geneworks, Adelaide, Australia), the bisulfi...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The present invention provides methods, compositions and kits for identifying the methylation status of a target nucleic acid molecule, the methylation status of a nucleotide locus in a target nucleic acid molecule, or for identifying A locus of one or more methylated or unmethylated nucleotides in a target nucleic acid molecule. The methylation state is achieved by treating the methylated target nucleic acid molecule with an agent that modifies one or more nucleotides in the target nucleic acid molecule as a function of the methylation state of the target nucleic acid molecule, methylation-specific amplification. Amplify the target nucleic acid molecule, fragment the amplification product, and detect one or more fragments to identify the methylation status of the target nucleic acid molecule, the methylation status of the nucleotide loci in the target nucleic acid molecule, or one or more of the target nucleic acid molecules loci of methylated or unmethylated nucleotides.

Description

[0001] related application [0002] This application claims priority to U.S. Provisional Patent Application No. 60 / 556,632, filed March 26, 2004, entitled "BASE SPECIFIC CLEAVAGE OFMETHYLATION-SPECIFIC AMPLIFICATION PRODUCTS INCOMBINATION WITH MASS ANALYSIS," to Mathias Ehrich and Dirk van den Boom, Its entire contents are hereby incorporated by reference. This application is subject to U.S. Application Serial No. 10 / 272,665 (to Andreas Braun, Christian Jurinke, and Dirk van den Boom, filed October 15, 2002, and entitled: "METHODS FOR GENERATING DATABASES AND DATABASES FOR IDENTIFYING POLYMORPHIC GENETIC MARKERS"; Involving U.S. Provisional Application Serial No. 10 / 723,365 (attorney docket number 2073) (to Dirk van den Boom and Sebastian , filed November 27, 2003, under the title "FRAGMENTATION-BASEDMETHODS AND SYSTEMS FOR SEQUENCE VARIATIONDETECTION AND DISCOVERY"); and relating to U.S. Provisional Series No. 60 / 466,006 (Attorney Docket No. P2070) (to Sebastian and Dirk va...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12P19/34C12Q1/68
CPCC12Q1/6848C12Q1/6858
Inventor 马蒂亚斯·埃里希迪克尔·约翰内斯·范德博姆
Owner SEQUENOM INC