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Quantifying method for oligonucleotide microarray

A technology of nucleic acid and target nucleic acid, which is applied in the system field of various nucleic acids, and can solve the problem of limited number of fluorescent dyes

Inactive Publication Date: 2009-11-25
HONEYWELL INT INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0009] However, real-time PCR technology has limitations, the most notable of which is that real-time PCR can only measure small quantities of nucleic acids in one reaction tube because the number of suitable fluorochromes with suitable corresponding fluorescence-emitting light sources is limited.

Method used

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  • Quantifying method for oligonucleotide microarray
  • Quantifying method for oligonucleotide microarray
  • Quantifying method for oligonucleotide microarray

Examples

Experimental program
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Embodiment 1

[0147] Quantified Microarray of Twelve Bacteria

[0148] The target panel included twelve different types of cells and two internal nucleic acid controls. Internal nucleic acid control A will be a known low concentration and internal nucleic acid control B will be a known high concentration. Primer sets included forward and reverse primers for all twelve bacterial species as well as internal nucleic acid controls A and B. The probe set will include probes for all twelve bacterial species as well as internal nucleic acid controls A and B. For example, target probe A is specific for target nucleic acid A, and target probe B is specific for target nucleic acid B.

[0149] Typically, there should be no cross-reactivity between probes for all twelve bacterial species and probes for internal nucleic acid controls A and B. Furthermore, typically there should be no cross-reactivity between the internal nucleic acid control probes A and B and the twelve target nucleic acids. Intern...

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Abstract

The invention relates to a quantifying method for an oligonucleotide microarray, and in particular provides a method and a device for simultaneously quantitatively measuring multiple nucleic acids from one or more pathogens in a sample in real time. Amplicons of target nucleic acids and internal nucleic acid contrast labeled by fluorescence are positioned on the matrix surface by hybridizing with target nucleic acid probes which are arranged in a predetermined two-dimensional mode and bound on the matrix surface and internal nucleic acid contrast probes. The hybridized amplicons can be detected by exciting fluorescence labels of the amplicons by using evanescent wave of proper wavelength light. By measuring fluorescence intensity on different positions of the matrix surface, abundance of hybridized amplicons of each target nucleic acid and the internal nucleic acid contrast can be determined.

Description

field of invention [0001] The present invention relates to systems and methods for quantitative measurement of nucleic acids, and more particularly to systems and methods for real-time, simultaneous quantitative analysis of multiple nucleic acids from one or more pathogens. Background of the invention [0002] Quantitative analysis of nucleic acids is very important in basic biological research and in fields such as clinical microbiology. Quantitative analysis is typically done in two stages. The target nucleic acid in the sample is first amplified to produce a detectable amount of the nucleic acid for use by the quantification tool. The detected amount of target nucleic acid is used to calculate the amount of nucleic acid originally present in the sample. [0003] Polymerase chain reaction (PCR) is a powerful method for amplifying nucleic acids, especially deoxyribonucleic acid (DNA). The key to practicing PCR is the use of thermostable DNA polymerases, proteins that cat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N21/64C12M1/34C12M1/00
CPCY02A50/30
Inventor Z·H·孙W·王Y·郑H·廖
Owner HONEYWELL INT INC
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