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A microarray system and a process for producing microarrays

A technology of microarrays and microbeads, applied in the field of microarrays, can solve the problems of complex preparation of microarrays, time-consuming, complex and expensive instruments, etc.

Active Publication Date: 2009-09-09
NAT UNIV OF SINGAPORE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These instruments are complex and expensive
All of the above factors make the preparation of microarrays more complex, more time consuming and thus more expensive to perform

Method used

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  • A microarray system and a process for producing microarrays
  • A microarray system and a process for producing microarrays
  • A microarray system and a process for producing microarrays

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0166] This example illustrates the use of bead-based microarrays for the detection of single nucleotide polymorphisms (SNPs).

[0167] Arrays of polyacrylamide gel pads were prepared on the surface of glass slides using ultraviolet photopolymerization. Each discrete gel pad is further composed of a square array of micropillars. Four sets of beads were prepared by separately conjugating the beads with oligonucleotide probes targeting specific SNPs. These four probes contain all possible single nucleotide substitutions of the ATGC at the target SNP site. Using a robotic dispenser, immobilize the first set of conjugated beads on designated gel pads using microtargeting. Loosely immobilized beads were washed away and the remaining immobilized beads were imaged using a CCD camera system. The positions of all immobilized beads are decoded from this image, and the data is stored in computer memory. This process was then repeated sequentially for a second set of conjugated beads ...

Embodiment 2

[0170] This example illustrates the use of bead-based microarrays for the detection of fecal indicator bacteria in environmental samples.

[0171] Twenty sets of probes were designed to target the 16S rRNA region of ten stool indicators. Each stool indicator is targeted by a pair of perfect match (PM) probes and a mismatch (MM) probe that differs by one single nucleotide in the central portion. Using the same method described in Example 1, ten sets of beads conjugated to PM probes were immobilized on different gel pads, and the position of the beads in each gel pad was characterized. This step is repeated for beads conjugated to MM probes, so that each gel pad contains beads conjugated to pairs of probes targeting a specific fecal indicator. These two groups of beads are distinguished based on their spatial codes.

[0172] Environmental samples suspected of containing fecal indicators were prepared and hybridized to immobilized beads. The presence of fecal indicators in unk...

Embodiment 3

[0174] This example demonstrates the performance of antibody-based assays using antibody-antigen interactions and DNA-based assays using DNA / DNA or DNA / RNA hybridization in parallel on the same chip.

[0175] Antibody preparation by biotinylation :

[0176] Antibodies (polyclonal mouse IgG and polyclonal mouse anti-IgG) were diluted to approximately 2 mg / ml. Biotin-NHS (10 mg / ml, dissolved in DMSO) was added to the antibody solution at a molar ratio of 1:1, 5:1 and 10:1 (M biotin:M antibody), and the reaction tube was placed at 25 Incubate overnight at °C. The biotinylated antibody was purified using a gel filtration column (D-salt dextran desalting column, Pierce.). Biotinylated antibodies were dissolved in PBS and stored at -20°C.

[0177] Method for FITC conjugation of antibodies :

[0178] Antibodies (polyclonal mouse IgG and polyclonal mouse anti-IgG, Arista Biological inc.) were diluted to approximately 2 mg / ml. FITC (10 mg / ml, dissolved in DMSO) was added to th...

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Abstract

The present invention provides a process for making a micro-array. The process comprises the step of depositing a population of microbeads on a substrate having at least one fiducial. The population being comprised of at least two sub-populations, preferably multiple sub-populations, each comprising a known active agent capable of specific binding with at least one target analyte. The said subpopulations are deposited sequentially and at discrete periods of each other. The process also comprises the step of making images of the substrate after deposition of each subpopulation. The images are then compared using the fiducial as a reference to thereby determine the location of each microbead and to identify the subpopulation, and its known active agent, based on differences between each image. Also disclosed in a system for using the microarray.

Description

technical field [0001] The present invention generally relates to microarrays for identifying the presence of target analytes in fluid samples. The invention also relates to methods for preparing microarrays. Background technique [0002] Microarrays are generally prepared by disposing biomolecules on a flat substrate. Known microarray preparation techniques can be divided into two methods: [0003] 1) the position of the biomolecule is specified prior to the transfer of the biomolecule to the substrate ("ordered biomolecule deposition"); and [0004] 2) The positions of the biomolecules are randomly distributed on the substrate ("random biomolecule deposition"). [0005] Ordered biomolecule deposition is by far the most common method used for microarray preparation. A detailed experimental description is given in Schena, M., "DNA Microarrays", Oxford University Press, 2001. A technical review is also given in Vivian G. et al., "Making and reading microarrays", Nature G...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/29C12Q1/68C07K17/14
CPCB01J2219/00722B01J2219/0074G06T7/0028C40B50/16B01J2219/00677C40B40/04B01J2219/00317B01J19/0046B01J2219/00459C40B50/18B01J2219/00648B01J2219/00659C12Q1/6834B01J2219/0054G01N21/6452C12Q1/6883B01J2219/00466C40B20/02B01J2219/00702C40B30/04G01N33/54313G06T5/50C40B60/14G06T7/0008G06T7/33C12Q2537/149C12Q2563/149C12Q2565/513C12Q2565/619C12Q1/6837G01N33/54373G01N33/545
Inventor 陶谛德刘文佐黄建国
Owner NAT UNIV OF SINGAPORE
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