KR101034273B1 - Method Of And System For Microarray Anaysis Using Magnetic Bead - Google Patents

Abstract

The present invention relates to a method and system for analyzing microarrays using magnetic beads. In one aspect of the present invention, a library preparation step of preparing a protein or cDNA or cRNA library to which magnetic beads are attached; A microarray hybridization step of hybridizing the prepared protein or cDNA or cRNA library to a previously prepared microarray; A magnetization step of magnetizing the magnetic beads on the hybridized microarray to read information; And a microarray analysis method using a magnetic bead comprising a; and a genetic analysis step of analyzing the result value read in the magnetization step is proposed.

The present invention enables much more economical microarray analysis than conventional microarray analysis using fluorescent materials.

Microarray, Library, Magnetic Bead, Magnetization, Digitization, Hard Disk Head

Description

Method and system for analyzing microarray using magnetic beads {Method Of And System For Microarray Anaysis Using Magnetic Bead}

The present invention relates to a method and system for analyzing microarrays using magnetic beads.

More specifically, the present invention relates to a method and system for analyzing microarrays using magnetic beads, in which magnetic beads, which are magnetic particles on a hybridized microarray, are magnetized and genetically analyzed by hybridizing proteins or cDNAs or cRNAs having magnetic beads attached thereto. .

Microarrays, such as DNA microarrays, are dense arrays of DNA molecules with known sequences on small substrates, such as slides. The advantage of microarrays is the ability to collectively explore large gene expression situations. DNA microarray technology is a fusion of various fields such as chemistry, molecular biology, mechanical engineering, and electronic engineering, and contributes greatly to genome-level research related to life phenomena. As a result of our decade of human genome business, we have deciphered all three billion DNA sequences that humans possess and obtained vast amounts of human genome DNA sequence information. The purpose of the human genome business is to decipher the sequence information of genomes and ultimately to reveal the function and structure of the genomes. Therefore, based on the vast sequence information, genome research is actively being conducted to derive biomedical useful information.

Because microarrays, such as DNA microarray technology, can put tens of thousands of gene fragments into a single microarray, it is possible to obtain information about the entire genome in a single experiment and to use only a few conventional genes. There is a characteristic that can get quite a lot of information that cannot be compared with the (Southern method) or Northern method (Northern method).

There are various types of microarrays, such as DNA chips, protein chips, cell chips, and nerve cell chips, depending on the type of material on the chip.

Looking at the basic principle of microarray, the part of the DNA base sequence that is converted into protein is transcribed into mRNA in the nucleus and exists as mRNA in the cell, and the amount of mRNA used to generate the protein through the translation process is the expression level of the gene. It can be used as a measure to indicate the relative expression level of genes using the complementary binding properties of DNA.

Conventional microarrays produce cDNAs or cRNAs with a fluorescent substance, ie, fluorescent dyes, which extract mRNA from tissues of interest, and hybridize them to pre-made DNA microarrays, where they have complementary sequences. Fluorescence appeared, and the relative expression levels of genes were determined by measuring the intensity of fluorescence.

The present invention is a method of expressing a gene in a manner using magnetic beads, such as computer hard disk head technology, by attaching magnetic beads to the extracted mRNA, instead of attaching a dye, that is, a fluorescent material. We propose a novel method and system for identifying and analyzing quantities.

The conventional technique of analyzing an image by attaching a fluorescent label is difficult to digitize, and has a disadvantage of separately purchasing an image analysis program package. In addition, a lot of time is required because a separate procedure is required to convert the analyzed image photograph back to digital. Most of all, the cost of conventional fluorescent labeling scanners is so high that they cannot be used in the home. Even if such a system is built, it will not be popularized at a high price.

The present invention proposes a microarray analysis method and system for detecting and analyzing the expression level of a gene in a magnetic method by attaching a magnetic bead to a protein, DNA, or RNA in order to solve this conventional problem. I would like to.

In order to achieve the above object, in one aspect of the present invention, a library preparation step of preparing a protein or cDNA or cRNA library attached magnetic beads (bead); A microarray hybridization step of hybridizing the prepared protein or cDNA or cRNA library to a prepared microarray; A magnetization step of magnetizing the magnetic beads on the hybridized microarray to read information; And a microarray analysis method using a magnetic bead comprising a; and a genetic analysis step of analyzing the result value read in the magnetization step is proposed.

In another preferred aspect of the present invention, in the aforementioned microarray analysis method, the genetic analysis step includes: a digitization step of digitizing the read value; And a digital analysis step of analyzing the digitized result value.

In a further preferred aspect to achieve the above object, in the above-described microarray analysis method, the magnetization step is characterized by magnetizing the magnetic beads on the hybridized microarray using a computer hard disk head method.

In another preferred aspect to achieve the above object, in the above-described microarray analysis method, the genetic analysis step, pre-processing the noise contained on the hybridized microarray prior to the genetic analysis of the result value Or further comprising a standardization step.

Also in a preferred aspect, the gene analysis step, after the above-described pre-processing or standardization step, further comprises a gene search step for finding genes that are expressed differently between a plurality of treatment groups. Or after the pre-processing or standardization step described above, further comprising a cluster analysis step for finding a cluster of genes having similar expression patterns.

In another preferred aspect to achieve the above object, in the above-described microarray analysis method, library preparation step, cDNA or cRNA synthesis step of synthesizing cDNA or cRNA by RNA extraction from a sample; And a labeling step of attaching and labeling the magnetic beads to the synthesized cDNA or cRNA.

Also in a preferred aspect, the labeling step, biotinylation step of preparing a biotin conjugates by attaching a biotin (Biotin) to the cDNA or cRNA; A magnetic bead complex preparation step of preparing a complex of magnetic beads with avidin or streptavidin or neutravidin; A magnetic bead binding step of hybridizing a biotin binder and a magnetic bead complex to bind magnetic beads to cDNA or cRNA by using a combination of biotin with avidin or streptavidin or neutravidin; And obtaining a bead labeling cDNA or cRNA to wash the biotin conjugate not bound to the magnetic bead complex to obtain a magnetic bead attached cDNA or cRNA.

In another aspect of the present invention for achieving the above object, a magnetic bead attached protein or cDNA or cRNA library module; A microarray hybridization unit for hybridizing the library module to a prepared microarray; A magnetic bead magnetizing unit for magnetizing the magnetic beads, which are magnetic materials on the microarray hybridized in the microarray hybridizing unit; And a genetic analyzer for reading information magnetized by the magnetic bead magnetizing unit and analyzing the read result value.

Also preferably, the above-described magnetic bead magnetization unit magnetizes the magnetic beads on the hybridized microarray using a computer hard disk head method.

According to the aspect of the present invention, a magnetic bead is attached to a magnetic bead such as a computer hard disk head technology by attaching a magnetic bead, rather than a method of attaching a dye, that is, a fluorescent substance, to a conventional extracted mRNA. This makes gene analysis much more economical.

In addition, it is easy to digitize, eliminates the need to purchase an image analysis program package, and saves time and economical microarray analysis by eliminating the need for a separate procedure to convert the analyzed image photograph back to digital. Became able

In addition, it is apparent that the detailed effects obtained according to the specific configuration according to the embodiments of the present invention can be clearly understood from the specific configuration of the embodiments to those skilled in the art even if not specifically described.

Hereinafter, preferred embodiments of the present invention for achieving the above object will be described with reference to the accompanying drawings. The same elements in the drawings will be described with the same reference numerals and symbols as much as possible even though they are shown in different drawings. In the following description of the present invention, when it is determined that a detailed description of a related known function or known configuration may unnecessarily obscure the subject matter of the present invention, the detailed description may be omitted or briefly described.

The present invention does not determine the relative expression level of genes by scanning fluorescent materials attached to conventional mRNAs, but by attaching magnetic beads to proteins or cDNAs or cRNAs, such as in computer hard disk heads. The method and system for analyzing the expression information of genes by magnetizing magnetic beads, which are magnetic materials, by using technology, and storing information or reading stored information.

First, a method of analyzing microarrays using magnetic beads, which is one of the present inventions, will be described.

Figure 1 shows a flow chart according to one embodiment of a microarray analysis method using a magnetic bead of the present invention, Figure 2 is another embodiment of a microarray analysis method using a magnetic bead of the present invention 3 shows a part of a flowchart according to another embodiment of a method of analyzing a microarray using magnetic beads, which is one of the present inventions, and FIG. 4 is a micro diagram using magnetic beads, which is one of the present inventions. A part of a flowchart according to another embodiment of an array analysis method is shown, and FIG. 5 is a part of a flowchart according to another embodiment of a microarray analysis method using magnetic beads, which is one of the present inventions. 6 is another embodiment of the microarray analysis method using magnetic beads of the present invention. It shows a portion of a flow chart according to one embodiment.

Referring to FIG. 1, one embodiment of a microarray analysis method using magnetic beads includes a library preparation step (S100), a microarray hybridization step (S200), a magnetization step (S300), and a genetic analysis step (S400). Is done.

First, in the library preparation step (S100), a protein or cDNA or cRNA library to which magnetic beads are attached is prepared.

Learn how to attach magnetic beads to proteins or DNA or RNA. To attach magnetic beads to proteins or DNA or RNA, first attach biotin and proteins or DNA or RNA, and attach magnetic beads to a medium such as avidin or streptavidin, and biotin attached protein material or genetic information. Avidin immobilized on the substance and magnetic beads is mixed to bind the magnetic beads with protein or DNA or RNA using a strong binding force between avidin and biotin. The unbound protein or DNA or RNA conjugate is washed away to prepare a magnetic bead attached protein or DNA or RNA.

In this way, a protein or cDNA or cRNA library to which magnetic beads are attached is prepared.

For example, the attachment of magnetic beads to proteins or DNA or RNA takes advantage of the binding properties of biotin and avidin.

The attachment of biotin to other molecules such as proteins is called biotinylation. Biotin is a water-soluble vitamin and is generally classified as a B-complex vitamin. Because the enzyme biotinidase catalyzes the biotinylation of histone, a DNA-binding protein, biotin is responsible for DNA replication and transcription. It plays an important role.

Biotin can be attached to a substance of interest, such as a protein or DNA or RNA. In the present invention, biotin is attached to a protein or DNA or RNA, more specifically to cDNA or cRNA. Specific attachment methods will be described later.

Preferably, as an example, referring to FIG. 5, the magnetic bead is attached to the cDNA or cRNA. Referring to FIG. 5, the library preparation step (S100) includes a cDNA or cRNA synthesis step (S110) and a labeling step (S120).

In the cDNA or cRNA synthesis step (S110) of FIG. 5, cDNA or cRNA is synthesized by extracting RNA from a target of interest, that is, a sample. Extracting RNA from a sample and cDNA or cRNA synthesis are only known techniques in the art, so a detailed description thereof will be omitted.

In the labeling step (S120), magnetic beads are attached to the synthesized cDNA or cRNA to be labeled. Attaching magnetic beads to cDNA or cRNA is a distinctive feature from attaching conventional dyes, ie fluorescent materials. In the present invention, gene expression information can be analyzed by reading information according to magnetization of magnetic beads attached to cDNA or cRNA.

More specific labeling methods are described in detail below.

Also in a more preferred embodiment, referring to Figure 6, the labeling step (S120) as shown in Figure 5, the labeling step, biotinylation step (S121), magnetic bead composite preparation step (S123), magnetic beads It comprises the binding step (S125) and the video labeling cDNA or cRNA acquisition step (S127). Although FIG. 6 describes a method of attaching magnetic beads to cDNA or cRNA, it may be obvious to those skilled in the art that such a method may be applied to a method of attaching magnetic beads to a protein.

In the biotinylation step (S121) as shown in FIG. 6, a biotin conjugate is prepared by attaching biotin to cDNA or cRNA. As described above, it is possible to prepare a biotin conjugate in which biotin is bound to a protein.

Specific examples of biotin and DNA or RNA attachment method, first, the terminal deoxynucleotidyl transferase (TdT) attaches TTP (UTP in the case of RNA) to the 3 'end of a single DNA. When the biotin-TTP complex is added and mixed with the TdT enzyme, the biotin is attached together with the TTP at the DNA 3 'end. Biotin-TdT here is made chemically cross-linked and commercially available.

These commercially available kits use TdT to promote nontemplate-directed nucleotide incorporation at the 3′-OH terminus of DNA. TdT represents the substrate affinity of single-stranded DNA. TdT, however, labels double DNA with 3 'overhang and blunt duplex, despite low efficiency. The biotin 3′-end DNA labeling kit is optimized for hybridizing biotin-11-dUTP (1-3 biotinylated ribonucleotides) at the 3 ′ end of the DNA strand. This labeling technique has the advantage of placing biotin at the 3 'end of a probe that is unlikely to interfere with sequence specific binding or hybridization of the protein. Biotin-attached DNA probes are non-isotope in a variety of applications, including electrophoretic mobility shift assays (EMSA), Northern or Southern blots, colony hybridization or in situ hybridization. It can be used to facilitate non-isotopic detection.

Next, referring to Figure 6 in the magnetic bead complex preparation step (S123) to prepare a complex of magnetic beads and avidin or streptavidin or neutravidin.

Magnetic bead complexes with avidin or streptavidin or neutravidin can be commercially purchased and chemically crosslinked with magnetic beads and avidin or streptavidin or neutravidin. -linking). Since it can be purchased commercially, a detailed description will be omitted.

Next, referring to FIG. 6, in the magnetic bead binding step (S125), the biotin conjugate is mixed with the magnetic bead complex to bind the magnetic beads to cDNA or cRNA by using a combination of biotin and avidin or streptavidin or neutravidin. . Although not shown, it is obvious to those skilled in the art to hybridize the magnetic beads to the protein by hybridizing the biotin conjugate and the magnetic bead complex associated with the protein. Hereinafter, the binding of the biotinylated DNA or RNA material to the magnetic bead complex will be described as an example.

Magnetic beads complexes of avidin or screptavidin or neutravidin immobilized on magnetic beads with biotinylated DNA or RNA material are mixed to take advantage of the strong binding force between avidin or screptavidin or neutravidin and biotin and Join the magnetic beads.

Displaced molecules such as avidin or screptavidin immobilized on magnetic beads are mixed and mixed with biotin conjugates. Beads complexes, such as avidin or streptavidin, are added to suspended hybridization buffers of biotin conjugates, and biotinylated molecules, such as DNA or RNA substances, are magnetic beads. Combined with.

Yeast or bacterial probes may be mixed with a hybrid buffer in which the conjugate of the prepared DNA or RNA and biotin and the magnetic bead complex are suspended.

DNA or RNA specimens are hybridized and magnetic beads attached DNA or RNA are obtained from the hybridized DNA or RNA specimens.

Hybrid buffers in which magnetic bead complexes are suspended can also be prepared by suspending magnetic bead complexes to form suspensions (suspensions) and slowly sucking and discarding the supernatant to create hybrid buffers in which magnetic beads are suspended. .

Biotin attached proteins or genetic information materials are used to capture avidin or streptavidin or neutravidin using a strong affinity of biotin with avidin or streptavidin or neutravidin. Any other protein that binds to the biotin attached molecule will stay with the beads. This is due to the binding between avidin and biotin. Hereinafter, the binding of biotin to avidin will be described based on the biotin protein conjugate.

Looking at the binding between avidin and biotin, avidin has the strongest binding force with biotin. Avidin shows a very strong affinity with biotin with a dissociation constant K _D of approximately ^10-15 / M, which is known as the strongest affinity between a protein and its ligand. In biochemical applications, streptavidin or neutravidin (NeutrAvidin), which bind very strongly with biotin, is often used in place of avidin.

Avidin's affinity for biotin is used for a wide range of biochemical assays, including specialty protein detection assays (western blots), Aenzyme-linked immunosorbent assays (ELIS), ELISPOTs, and pull-down assays. . Avidin immobilized on a rigid support is also used as a purification medium to capture biotinylated proteins or nucleic acid molecules. For example, cell surface proteins may be specifically attached to membrane impermeable biotin responders, in which case they may be specifically captured using avidin based supports.

In addition, referring to FIG. 6, in the bead labeling cDNA or cRNA acquisition step (S127), the biotin conjugate that is not bound to the magnetic bead complex is washed to obtain a magnetic bead attached cDNA or cRNA. Although not shown, it will be apparent to those skilled in the art that a non-binding biotin protein conjugate may be washed away to obtain a magnetic bead attached protein during the binding process of the biotin protein conjugate to the magnetic bead complex. Can be.

Beads complexes, such as avidin or streptavidin, are added to suspended hybridization buffers of biotin conjugates, and biotinylated molecules, such as DNA or RNA, are magnetic It will bind to the beads, and the other biotin binding materials that are not bound to the magnetic bead complex can then be washed away.

Unbound biotin conjugates are washed away to obtain magnetic bead attached cDNA or cRNA.

Next, referring to FIG. 1, in the microarray hybridization step (S200), the prepared protein or cDNA or cRNA is hybridized to a previously prepared microarray.

Microarrays prepared in advance may be manufactured by various methods. For example, it can generally be classified into four types according to the manufacturing method, such as a pin microarray, an inkjet printing array, an optical etching chip, an electronic array, and the like. Pin microarray is a method of taking a gene from a well plate containing a gene amplified by PCR and directing it to a microarray slide. An inkjet method is performed by using a cartridge such as an inkjet printer containing a gene. Genetic spraying genes on microarray slides by optical force, photoetching chip is a method of manufacturing chips by directly synthesizing bases by applying photoetching technology used in semiconductor chip fabrication, and electron array is a negative (-) This is a method of attaching a desired gene using charge properties.

Microarrays are also classified according to the type of probe attached to the chip. Probe refers to an individual gene or EST attached to a chip. Depending on the type of probe, there are protein chips, DNA chips, and the like, and DNA chips may be classified into, for example, cDNA or oligonucleotide chips.

Hybridization of proteins, cDNAs, or cRNAs with magnetic beads attached to pre-made microarrays enables magnetization of magnetic bodies in regions having complementary sequences to identify magnetization information.

Referring to FIG. 1, in the magnetization step (S300), which is the next step of the microarray analysis method using magnetic beads, magnetized magnetic beads on a hybridized microarray are magnetized to read information.

Preferably, as another embodiment, in the magnetization step (S300) to magnetize the magnetic beads on the hybridized microarray using a computer hard disk head method. The head of a computer hard disk magnetizes a magnetic substance coated on a platter surface to store information and read stored information. By applying this technique, the magnetic beads on a microarray are magnetized to read magnetized information. A computer hard disk head type magnetization method can be obvious to those skilled in the art.

8 is a conceptual diagram of a microarray analysis using magnetic beads according to one of the present invention.

The microarray analysis method according to the present invention is similar to the conventional microarray analysis method, but instead of attaching a fluorescent material to the cDNA, magnetic beads, preferred embodiments iron bead (iron bead) by attaching the amount of RNA expressed for each gene Since it can be identified as a magnetic head of the hard disk, it can be automatically digitized, and accordingly, it can be stored in a database in real time without undergoing a separate image conversion and scoring process according to each expressed RNA amount.

Table 1 below shows one embodiment of an information table which receives magnetization information of magnetic beads on a microarray as a computer hard disk head.

[Table 1] Information table accepted as magnetic head

In Table 1, a small rectangle of nine numbers is called a feature. This is where the same gene is attached. If the conventional fluorescence labeling method is used, the intensity of fluorescence increases in order from the sum of nine numbers. "0" is a portion that is not hybridized with the cDNA attached to the iron bead (Iron bead), "1" means that it is hybridized with the cDNA.

It is possible to score genes expressed by the sum of the numbers according to the number of tests for each gene, and the following Table 2 shows an example of a scoring table of expressed genes.

TABLE 2 Scoring Table of Expressed Genes

Number of times 1 time Episode 2 3rd time 4 times Sum a gene 3 0 One 2 6 b gene 2 2 3 2 9 c gene 6 6 2 4 18 d gene 8 7 6 8 29 e gene One 0 2 One 4 f gene 2 3 3 4 12 g gene 5 4 8 6 23

By developing the microarray analysis method and system according to the present invention, it is possible to produce a scanner capable of microarray analysis at a low price by applying the current advanced hard disk production technology without using an expensive fluorescence analysis scanner.

In addition, referring to FIG. 1, in the genetic analysis step S400, which is the last step of the microarray analysis method using magnetic beads, the resultant value read in the magnetization step S300 described above is analyzed. The magnetized information is preferably digitized and read, and the digital information value is analyzed. For example, according to the general structure of the cDNA microarray data, it can be expressed and analyzed as a row representing a gene, a column with sample information, and a digitized expression value.

Preferably, as another embodiment, referring to FIG. 2, the genetic analysis step S400 includes a digitization step S410 and a digital analysis step S420.

In the digitization step S410, the value read in the magnetization step S300 is digitized, and in the digital analysis step S420, the digitized result value is analyzed.

According to the embodiment of the present invention, instead of attaching a fluorescent material to the cDNA, magnetic beads, preferably iron beads (Iron bead) by attaching the amount of RNA expressed by each gene is automatically identified as the magnetic head of the hard disk Digitized. Therefore, there is an advantage that can be stored in the database in real time without going through a separate image conversion and scoring process according to each expressed RNA amount.

Also, in another embodiment, referring to FIG. 3, in the microarray analysis methods, the genetic analysis step S400 may be performed by processing noise included in the hybridized microarray prior to genetic analysis of the resultant value. It further comprises a pre-processing or standardization step (S399). This pretreatment or standardization step (S399) is to adjust the various types of variations that occur in the microarray experiment. Microarray data contain more noise or non-biological variation than normal experimental data, and often show a pattern in the data. In particular, since noise with a certain pattern may have a fatal effect on the analysis result, it is standardized to remove noise by preparing a pretreatment process in analyzing DNA microarray data or considering noise in the analysis. As a standardization method, a method of systematically removing mutations using a statistical regression model is widely used.

Also, more preferably, referring to FIG. 4, in the microarray analysis method, the gene analysis step S400 may be performed after the pretreatment or standardization step S399 as shown in FIG. 3. It further comprises a cluster analysis step (S401b).

In the gene search step (S401a) to find a gene that is expressed differently between a plurality of treatment groups. As an example of finding genes that are expressed differently between groups, when a chip experiment using cancer tissues and normal tissues is performed, genes that are strongly expressed in cancer tissues can be found compared to normal tissues. The genes that are related can be identified.

In the cluster analysis step (S401b), a cluster of genes having similar expression patterns is found. Cluster analysis can reveal the function of a gene whose function is unknown.

It looks at the microarray analysis system using a magnetic bead of another one of the present invention. Description of the same functions and operations as described in the microarray analysis method using magnetic beads will be omitted.

Figure 7 is a block diagram showing one embodiment of a microarray analysis system using magnetic beads, which is another of the present invention.

Referring to FIG. 7, one embodiment of the microarray analysis system includes a bead-attached library module 100, a microarray hybridization unit 200, a magnetic bead magnetization unit 300, and a genetic analysis unit 400. Is done.

The beaded library module 100 is a magnetic bead attached protein or cDNA or cRNA library, and magnetic bead attachment to the protein or cDNA or cRNA is described above in the Examples of Microarray Assay.

The microarray hybridization unit 200 hybridizes the library module 100 to a prepared microarray. Microarray hybridization refers to what has been described in the examples of the microarray analysis method described above.

The magnetic bead magnetizing unit 300 magnetizes magnetic beads, which are magnetic bodies on the microarray hybridized by the microarray hybridization unit 200.

Preferably, the magnetic bead magnetization unit 300 magnetizes the magnetic beads on the hybridized microarray using a computer hard disk head method.

The genetic analyzer 400 reads the information magnetized by the magnetic bead magnetizer 300 and analyzes the read result value.

The magnetized information is preferably digitized and read, and the digital information value is analyzed.

In the above, with reference to the accompanying drawings, the present invention has been described with reference to the preferred embodiments. Although specific terms have been used herein, they are used only for the purpose of describing the present invention and are not used to limit the scope of the present invention as defined in the meaning or claims. In addition, the accompanying drawings and the foregoing embodiments are described by way of example to help those skilled in the art to understand the present invention. Therefore, it will be apparent to those skilled in the art that the present invention may be implemented in a modified form without departing from the essential characteristics of the present invention. The foregoing embodiments are, therefore, to be regarded in an illustrative rather than a restrictive sense, and the scope of the present invention should be construed in accordance with the invention set forth in the appended claims rather than the foregoing embodiments, the scope of which is usually defined in the art. It includes various modifications, alternatives, and equivalents by those with knowledge of.

Figure 1 shows a flow chart according to one embodiment of a microarray analysis method using a magnetic bead of the present invention,

Figure 2 shows a part of a flow chart according to another embodiment of a microarray analysis method using a magnetic bead of the present invention,

Figure 3 shows a part of a flow chart according to another embodiment of a microarray analysis method using a magnetic bead of the present invention,

Figure 4 shows a part of a flow chart according to another embodiment of a microarray analysis method using a magnetic bead of the present invention,

Figure 5 shows a part of a flow chart according to another embodiment of a microarray analysis method using a magnetic bead of the present invention,

Figure 6 shows a part of a flow chart according to another embodiment of a microarray analysis method using a magnetic bead of the present invention,

7 is a block diagram showing one embodiment of a microarray analysis system using magnetic beads, which is another of the present invention;

8 is a conceptual diagram of a microarray analysis using magnetic beads according to one of the present invention.

<Description of reference numerals for the main parts of the drawings>

100: bead attached library module 200: microarray hybridization unit

300: magnetic bead magnetization unit 400: gene analysis unit

Claims (9)

A library preparation step of preparing a protein or cDNA or cRNA library to which magnetic beads are attached; A microarray hybridization step of hybridizing the prepared protein or cDNA or cRNA library to a microarray prepared in advance; A magnetization step of magnetizing the magnetic beads on the hybridized microarray to read information; And Genetic analysis step of analyzing the result value read in the magnetization step; Microarray analysis method using a magnetic bead comprising a. The method according to claim 1, wherein the genetic analysis step: Digitizing the digitized value; And And a digital analysis step of analyzing the digitized result value. The method according to claim 1, The magnetizing step is a microarray analysis method using magnetic beads, characterized in that the magnetized magnetic beads on the hybridized microarray using a computer hard disk head method. The method according to any one of claims 1 to 3, wherein the genetic analysis step, And a pre-processing or normalizing step of processing noise included in the hybridized microarray prior to the genetic analysis of the resultant value. The method according to claim 4, wherein the genetic analysis step, After the pretreatment or normalization step, a gene search step for finding genes that are expressed differently among a plurality of treatment groups, or Microarray analysis method using magnetic beads, characterized in that further comprises a cluster analysis step for finding a cluster of similar genes. The method of claim 1, wherein the library preparation step is: CDNA or cRNA synthesis step of synthesizing cDNA or cRNA by RNA extraction from the sample; And Labeling step of attaching and labeling the magnetic beads on the synthesized cDNA or cRNA; Microarray analysis method using a magnetic bead comprising the. The method of claim 6, wherein the labeling step: biotinylation step of preparing biotin conjugates by attaching biotin to cDNA or cRNA; A magnetic bead complex preparation step of preparing a complex of magnetic beads with avidin or streptavidin or neutravidin; A magnetic bead binding step of hybridizing a biotin binder and a magnetic bead complex to bind magnetic beads to cDNA or cRNA by using a combination of biotin with avidin or streptavidin or neutravidin; And Obtaining a bead-labeled cDNA or cRNA to wash off the biotin conjugates not bound to the magnetic bead complex to obtain a magnetic bead attached cDNA or cRNA; Microarray analysis method using magnetic beads, characterized in that comprises a. Magnetic bead attached proteins or cDNA or cRNA library modules; A microarray hybridization unit for hybridizing the library module to a prepared microarray; A magnetic bead magnetizing unit for magnetizing magnetic beads, which are magnetic bodies on the microarray hybridized by the microarray hybridizing unit; And And a genetic analyzer for reading information magnetized by the magnetic bead magnetizer and analyzing the read result value. The method according to claim 8, The magnetic bead magnetizing unit magnetizing the magnetic beads on the hybridized microarray using a computer hard disk head method, the microarray analysis system using magnetic beads.

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