CN112858693A - Biomolecule detection method - Google Patents
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
- G01N33/6845—Methods of identifying protein-protein interactions in protein mixtures
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
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- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/5308—Immunoassay; Biospecific binding assay; Materials therefor for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
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Abstract
The invention discloses a biomolecule detection method, and relates to the technical field of biomolecule detection. The detection method comprises protein in-vitro combination test detection, co-immunoprecipitation detection and gene detection; the protein in-vitro binding test detection method comprises the following steps of protein-protein interaction research, RNA/DNA binding protein interaction research and RNA/DNA interaction research and protein-protein interaction research detection, wherein the Step 01: preparing a relevant detection instrument comprising a biomolecule interaction analyzer; step 02: constructing a GST-target protein fusion expression vector; step 03: MS detecting the expression of the fusion protein; step 04: obtaining total protein lysate. The biomolecule detection method can detect a complex formed in vivo, eliminates false positive brought by target protein, and simultaneously, endogenous target protein is processed, modified and mature protein, so that interaction of the modified protein can be detected, and the detection completion degree is higher.
Description
Technical Field
The invention relates to the technical field of biomolecule detection, in particular to a biomolecule detection method.
Background
Biomolecules generally refer to various molecules specific to organisms, all of which are organic matters, and generally refer to various molecules specific to organisms, all of which are organic matters, typical cells contain ten thousand to one hundred thousand biomolecules, wherein nearly half of the biomolecules are small molecules, the molecular weight is generally below 500, the rest are polymers of biological small molecules, the molecular weight is very large, generally more than ten thousand, and some are as high as 10 ten thousand, so that the biomolecules are called as biological macromolecules, small molecular units of the biological macromolecules are formed, the biomacromolecules are called as members, and amino acids, nucleotides and monosaccharides are members forming proteins, nucleic acids and polysaccharides respectively; the biological molecules have own specific structures, the molecular weight of the biological macromolecules is large, the components are various in types and large in quantity, the arrangement sequence is diversified, so that the structure is very complex, only 10 proteins are estimated, the biological molecules are ordered, each biological molecule has own structural characteristics, and all the biological molecules exist in a life system in a certain order (organization);
molecular diagnostics refers to techniques for making a diagnosis by detecting changes in the structure or expression level of genetic material in a patient using molecular biology methods. Molecular diagnosis is a main method for prediction diagnosis, and can be used for diagnosis of individual genetic diseases and prenatal diagnosis. Molecular diagnosis mainly refers to the detection of various structural proteins, enzymes, antigen antibodies and immune active molecular genes related to coding diseases, and the main technologies of molecular diagnosis include nucleic acid molecular hybridization, polymerase chain reaction and biochip technology. 1. The nucleic acid molecule hybridization technique has a process of association of a certain complementary sequence and a nucleotide single strand into a heteroduplex in a liquid phase or a solid phase according to the base complementary pairing principle, and is called nucleic acid molecule hybridization. The two hybridized parts are the nucleic acid sequence to be detected and the probe sequence. The technology can be used for qualitatively or quantitatively detecting specific DNA or RNA sequences. 2. Polymerase chain reaction, principle: PCR is that template DNA, primer and four kinds of deoxyribose nucleic acid triphosphate (dNTP) take place enzymatic polymerization reaction under the action of DNA polymerase, amplify out the DNA of the necessary purpose; comprises three basic steps: heating and denaturing the double-stranded DNA template into single strands (denaturation); complementary pairing (annealing) of primers to single-stranded DNA at low temperature; extending the primer along the template DNA catalyzed by TapDNA polymerase at a suitable temperature; 3. the biochip technology is a nucleic acid analysis and detection technology combining molecular biology and microelectronic technology which is developed in recent years; the primary goal of biochip technology is for DNA sequencing, gene expression profiling and gene mutant detection and analysis, and is also known as DNA chip or gene chip technology; because the technology is expanded to the non-nucleic acid fields of immune reaction, receptor combination and the like, and a protein chip, an immune chip, a cell chip, a tissue chip and the like appear, the technology of the biochip is called to be more in line with the development trend; the development of molecular diagnosis technology is highly emphasized in all countries in the world, and gene chips will become the mainstream of the development of a new generation of molecular diagnosis reagent. The gene chip is a crystal combined with multiple disciplines of molecular biology, microelectronics, computers and the like, integrates various modern high-precision technologies, and is praised as a final product of the diagnosis industry by experts. The gene chip has the function of simultaneously detecting a plurality of target points and has the characteristics of rapidness and effectiveness;
the current biomolecule detection method has the disadvantages of insufficient accuracy, complex operation steps, long detection time and insufficient sensitivity, and meanwhile, the traditional detection method has poor adaptability in a single detection mode and is difficult to adapt to biomolecule detection in multiple fields; to this end, we propose a method for biomolecule detection.
Disclosure of Invention
The present invention has been made in an effort to solve the above problems occurring in the background, and an object of the present invention is to provide a method for detecting a biomolecule.
In order to solve the technical problems, the invention is realized by the following technical scheme:
the invention relates to a biomolecule detection method, which comprises protein in-vitro combination test detection, co-immunoprecipitation detection and gene detection;
the protein in-vitro binding test detection comprises the study of protein-protein interaction and the study of RNA/DNA binding protein interaction with target RNA/DNA thereof, and the protein-protein interaction study detection method comprises the following steps:
step 01: preparing a relevant detection instrument comprising a biomolecule interaction analyzer;
step 02: constructing a GST-target protein fusion expression vector;
step 03: MS detecting the expression of the fusion protein;
step 04: obtaining total protein lysate;
step 05: using a biomolecule interaction analyzer to perform a protein in-vitro binding experiment to obtain interaction binding protein;
step 06: purifying and analyzing the binding protein to obtain a detection result;
step 07: and cleaning the experimental detection instrument and cleaning the detection environment.
The interaction research and detection method between the RNA/DNA binding protein and the target RNA/DNA thereof comprises the following steps:
step 11: preparing a relevant detection instrument comprising a biomolecule interaction analyzer;
step 12: preparing an RNA/DNA biotin probe;
step 13: preparing cell lysate;
step 14: obtaining total protein lysate;
step 15: RNA/DNA Pull down regulates the interaction binding protein;
step 16: purifying and analyzing the binding protein to obtain a detection result;
step 17: and cleaning the experimental detection instrument and cleaning the detection environment.
The co-immunoprecipitation detection method comprises the following steps:
step 21: taking a target protein endogenous to the cell as a bait, and performing co-incubation on the target protein antibody and the total cell protein;
step 22: forming an immune complex based on Step 21;
step 23: adding protein A/G capable of being combined with the target protein antibody into the immune complex to form a 'combined protein-target protein antibody' complex;
step 24: purifying the immune complex and then carrying out gel electrophoresis separation;
step 25: based on Step24, the protein is purified and the antigen or antibody is detected by combining Western blot or mass spectrometry.
When the co-immunoprecipitation detection is carried out, samples such as tissues, cells, protein solution and the like need to be prepared, and related information of the protein to be detected is needed; meanwhile, if an overexpression vector of the gene needs to be constructed, the overexpression vector and related information thereof also need to be provided; and other immunoprecipitation related information.
The gene detection method comprises probe method detection, MassARRAY method detection and Illumina BeadXpress method detection, and the detection method of the probe method detection comprises the following steps:
step 31: extracting DNA of a related detection object;
step 32: performing sample quality detection on the extracted DNA;
step 33: designing and synthesizing a detection probe;
step 34: carrying out a preliminary experiment to obtain corresponding experiment basic data;
step 35: detecting by a fluorescent quantitative PCR instrument;
step 36: and (5) completing detection, and analyzing data to obtain a detection result.
The probe method can directly read the genotype (homozygote XX or YY, heterozygote XY) of the SNP in each sample in the experimental result, does not need to analyze the sequence, and has intuition.
The detection method for the MassARRAY method comprises the following steps:
step 41: preparing a detection sample;
step 42: carrying out the integrity detection of the sample and the pretreatment of the sample;
step 43: designing and synthesizing a primer;
step 44: performing PCR amplification, purification, extension or transcriptional cleavage based on Step 43;
step 45: performing spotting and mass spectrometry on the detection sample finished in the Step 44;
step 46: and obtaining a detection result.
The detection method for detecting the Illumina BeadXpress method comprises the following steps:
step 51: customizing a gene chip;
step 52: preparing a gene sample, and performing quality detection on the prepared sample;
step 53: carrying out on-machine detection on the sample by using a detection instrument;
step 54: analyzing the obtained detection data;
step 55: and obtaining a detection result.
The bead chip has the characteristics of high density, high repeatability, high sensitivity, low sample loading amount, flexible customization and the like, and has extremely high integration density, so that extremely high detection and screening speed is obtained, and the cost can be obviously reduced during high-throughput screening.
Preferably, the protein in vitro binding assay is a method for detecting protein-protein interaction under in vitro conditions, and the corresponding target protein is obtained by SDS-page electrophoretic analysis, MS detection, verification of protein-protein interaction or screening.
Preferably, the protein in vitro binding assay is an in vitro validation/search for interacting proteins using a labeled antibody for detection.
Preferably, the protein in vitro binding experiment biotin-labeled RNA/DNA probe is incubated with a cytoplasmic protein extract to form an RNA/DNA-protein complex.
Preferably, the co-immunoprecipitation technique is a method for studying protein interactions based on the interaction between an antibody and an antigen.
Preferably, the probe method detection is based on fluorescent quantitative PCR, and PCR primers and PCR amplification are respectively designed aiming at different SNP sites on a chromosome.
Preferably, the MassARRAY method detects DNA molecular fragments excited by laser, and then determines the molecular weight of the fragments by flight time.
Preferably, the Illumina BeadXpress method detection is suitable for high-throughput detection, and can be used for simultaneously detecting a plurality of SNP sites for genome chip result confirmation.
The invention has the following beneficial effects:
when the biomolecule detection method is used for co-immunoprecipitation detection, the co-immunoprecipitation is used for detecting the interaction between proteins under physiological conditions, so that not only can a complex formed in vivo be detected, but also false positive caused by overexpression of the target protein can be eliminated, and next, endogenous target protein is processed, modified and mature protein, so that the interaction of the modified protein can be detected, and the detection completion degree is higher.
In the biomolecule detection method, the gene detection is based on the polymorphism of a DNA sequence caused by the mutation of a single nucleotide in a chromosome genome, exists in the forms of transversion, conversion, insertion, deletion and the like of a single base, is used as a third-generation molecular marker, has great development potential, is widely applied to a plurality of fields of biology, agriculture, medicine, biological evolution and the like, and plays an important role in the aspects of molecular genetics, pharmacogenetics, forensic medicine, diagnosis and treatment of diseases and the like.
In the biomolecule detection method, the probe method is usually used for analyzing a small amount of SNP sites, and has the advantages of low cost and high speed.
Of course, it is not necessary for any product in which the invention is practiced to achieve all of the above-described advantages at the same time.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art that other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a block diagram of the operation steps of a biomolecule detection method according to the present invention;
FIG. 2 is a flowchart of a protein-protein interaction assay of a biomolecule assay of the present invention;
FIG. 3 is a flow chart of the method for detecting the interaction between RNA/DNA binding protein and its target RNA/DNA according to the method for detecting biomolecules of the present invention;
FIG. 4 is a flow chart of the co-immunoprecipitation assay of the biomolecule assay of the present invention;
FIG. 5 is a flow chart of a probe-based detection method of the biomolecule detection method of the present invention;
FIG. 6 is a flowchart of the detection method of the MassARRAY method of the biomolecule detection method of the present invention;
FIG. 7 is a flow chart of the detection method of Illumina BeadXpress method of the biomolecule detection method of the present invention;
FIG. 8 is a schematic diagram of the working principle of the interaction between RNA/DNA binding protein and its target RNA/DNA for researching and detecting the present invention;
FIG. 9 is a schematic diagram of the working principle of co-immunoprecipitation detection according to the biomolecule detection method of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Please refer to fig. 1-2: the invention relates to a biomolecule detection method, which comprises protein in vitro combination test detection, co-immunoprecipitation detection and gene detection;
the protein in vitro binding test detection method comprises the following steps of protein-protein interaction research and RNA/DNA binding protein and target RNA/DNA interaction research, and the protein-protein interaction research detection method comprises the following steps:
step 01: preparing a relevant detection instrument comprising a biomolecule interaction analyzer;
step 02: constructing a GST-target protein fusion expression vector;
step 03: MS detecting the expression of the fusion protein;
step 04: obtaining total protein lysate;
step 05: using a biomolecule interaction analyzer to perform a protein in-vitro binding experiment to obtain interaction binding protein;
step 06: purifying and analyzing the binding protein to obtain a detection result;
step 07: and cleaning the experimental detection instrument and cleaning the detection environment.
Please refer to fig. 3 and 8: the interaction research and detection method between the RNA/DNA binding protein and the target RNA/DNA thereof comprises the following steps:
step 11: preparing a relevant detection instrument comprising a biomolecule interaction analyzer;
step 12: preparing an RNA/DNA biotin probe;
step 13: preparing cell lysate;
step 14: obtaining total protein lysate;
step 15: RNA/DNA Pull down regulates the interaction binding protein;
step 16: purifying and analyzing the binding protein to obtain a detection result;
step 17: and cleaning the experimental detection instrument and cleaning the detection environment.
Please refer to fig. 4 and 9: the co-immunoprecipitation detection method comprises the following steps:
step 21: taking a target protein endogenous to the cell as a bait, and performing co-incubation on the target protein antibody and the total cell protein;
step 22: forming an immune complex based on Step 21;
step 23: adding protein A/G capable of being combined with the target protein antibody into the immune complex to form a 'combined protein-target protein antibody' complex;
step 24: purifying the immune complex and then carrying out gel electrophoresis separation;
step 25: based on Step24, the protein is purified and the antigen or antibody is detected by combining Western blot or mass spectrometry.
Please refer to fig. 5: the gene detection method comprises the following steps of probe method detection, MassARRAY method detection and Illumina BeadXpress method detection, and the detection method of the probe method detection comprises the following steps:
step 31: extracting DNA of a related detection object;
step 32: performing sample quality detection on the extracted DNA;
step 33: designing and synthesizing a detection probe;
step 34: carrying out a preliminary experiment to obtain corresponding experiment basic data;
step 35: detecting by a fluorescent quantitative PCR instrument;
step 36: and (5) completing detection, and analyzing data to obtain a detection result.
The probe method can directly read the genotype (homozygote XX or YY, heterozygote XY) of the SNP in each sample in the experimental result, does not need to analyze the sequence, and has intuition.
Please refer to fig. 6: the detection method of the MassARRAY method comprises the following steps:
step 41: preparing a detection sample;
step 42: carrying out the integrity detection of the sample and the pretreatment of the sample;
step 43: designing and synthesizing a primer;
step 44: performing PCR amplification, purification, extension or transcriptional cleavage based on Step 43;
step 45: performing spotting and mass spectrometry on the detection sample finished in the Step 44;
step 46: and obtaining a detection result.
The MassARRAY method has flexible experimental design, high accuracy of typing results and high cost performance, can detect hundreds to thousands of samples, and has sample flux from hundreds to tens of thousands.
Please refer to fig. 7: the detection method for detecting the Illumina BeadXpress method comprises the following steps:
step 51: customizing a gene chip;
step 52: preparing a gene sample, and performing quality detection on the prepared sample;
step 53: carrying out on-machine detection on the sample by using a detection instrument;
step 54: analyzing the obtained detection data;
step 55: and obtaining a detection result.
The bead chip has the characteristics of high density, high repeatability, high sensitivity, low sample loading amount, flexible customization and the like, and has extremely high integration density, so that extremely high detection and screening speed is obtained, and the cost can be obviously reduced during high-throughput screening.
The protein in-vitro combination experiment is a method for detecting the interaction between proteins under in-vitro conditions, and the corresponding target protein is obtained by SDS-page electrophoretic analysis, MS detection and verification of the interaction between the proteins or screening.
Wherein, the protein in vitro combination experiment is a technology for verifying/searching interacting protein in vitro, and a label antibody is adopted for detection.
Wherein, the protein in vitro combination experiment biotin labeled RNA/DNA probe is incubated with the cytoplasmic protein extract to form an RNA/DNA-protein complex.
Among them, the co-immunoprecipitation technology is a method for studying the interaction of proteins based on the action between an antibody and an antigen.
Wherein, the probe method detection is based on fluorescence quantitative PCR, and PCR primers and PCR amplification are respectively designed aiming at different SNP sites on a chromosome.
The MassARRAY method detects DNA molecular fragments excited by laser, and then judges the molecular weight of the fragments by flight time.
The Illumina BeadXpress method is suitable for high-throughput detection, can detect a plurality of SNP sites simultaneously, and is used for confirming the result of a genome chip.
In the invention, RNA/DNA Pull down is one of the main means for detecting the interaction between RNA/DNA binding protein and target RNA/DNA thereof, RNA/DNA probe is labeled by biotin and incubated with cytoplasmic protein extract to form RNA/DNA-protein complex, the complex can be combined with magnetic beads labeled by streptavidin so as to be separated from other components in the incubation liquid, and after the complex is eluted, whether specific RNA/DNA binding protein interacts with RNA/DNA or not can be detected, and the protein type can be identified by Mass Spectrometry (MS) on the protein.
In the invention, when the co-immunoprecipitation detection is carried out, samples such as tissues, cells, protein solution and the like need to be prepared, and the related information of the protein to be detected; meanwhile, if an overexpression vector of the gene needs to be constructed, the overexpression vector and related information thereof also need to be provided; and other immunoprecipitation related information.
In the invention, the probe method can directly read the genotype (homozygote XX or YY, heterozygote XY) of SNP in each sample in the experimental result, does not need to analyze the sequence, and has intuition; the MassARRAY method has flexible experimental design, high accuracy of typing results and high cost performance, can detect hundreds to thousands of samples, and has sample flux from hundreds to tens of thousands; the Illumina bead Xpress method is suitable for high-throughput detection, and the microbead chip has the characteristics of high density, high repeatability, high sensitivity, low sample loading amount, flexibility in customization and the like, and has extremely high integration density, so that extremely high detection and screening speed is obtained, and the cost can be obviously reduced during high-throughput screening.
In the invention, the Gene detection method also comprises a SNaPshot method, and the principle is that the fluorescence BigDye sequencing of only one basic group is prolonged.
In the description herein, references to the description of "one embodiment," "an example," "a specific example" or the like are intended to mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, the schematic representations of the terms used above do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
The preferred embodiments of the invention disclosed above are intended to be illustrative only. The preferred embodiments are not intended to be exhaustive or to limit the invention to the precise embodiments disclosed. Obviously, many modifications and variations are possible in light of the above teaching. The embodiments were chosen and described in order to best explain the principles of the invention and the practical application, to thereby enable others skilled in the art to best utilize the invention. The invention is limited only by the claims and their full scope and equivalents.
Claims (8)
1. A biomolecule detection method, comprising: the detection method comprises protein in-vitro binding test detection, co-immunoprecipitation detection and gene detection;
the protein in-vitro binding test detection comprises the study of protein-protein interaction and the study of RNA/DNA binding protein interaction with target RNA/DNA thereof, and the protein-protein interaction study detection method comprises the following steps:
step 01: preparing a relevant detection instrument comprising a biomolecule interaction analyzer;
step 02: constructing a GST-target protein fusion expression vector;
step 03: MS detecting the expression of the fusion protein;
step 04: obtaining total protein lysate;
step 05: using a biomolecule interaction analyzer to perform a protein in-vitro binding experiment to obtain interaction binding protein;
step 06: purifying and analyzing the binding protein to obtain a detection result;
step 07: and cleaning the experimental detection instrument and cleaning the detection environment.
The interaction research and detection method between the RNA/DNA binding protein and the target RNA/DNA thereof comprises the following steps:
step 11: preparing a relevant detection instrument comprising a biomolecule interaction analyzer;
step 12: preparing an RNA/DNA biotin probe;
step 13: preparing cell lysate;
step 14: obtaining total protein lysate;
step 15: RNA/DNA Pull down regulates the interaction binding protein;
step 16: purifying and analyzing the binding protein to obtain a detection result;
step 17: and cleaning the experimental detection instrument and cleaning the detection environment.
The co-immunoprecipitation detection method comprises the following steps:
step 21: taking a target protein endogenous to the cell as a bait, and performing co-incubation on the target protein antibody and the total cell protein;
step 22: forming an immune complex based on Step 21;
step 23: adding protein A/G capable of being combined with the target protein antibody into the immune complex to form a 'combined protein-target protein antibody' complex;
step 24: purifying the immune complex and then carrying out gel electrophoresis separation;
step 25: based on Step24, the protein is purified and the antigen or antibody is detected by combining Western blot or mass spectrometry.
The gene detection method comprises probe method detection, MassARRAY method detection and Illumina BeadXpress method detection, and the detection method of the probe method detection comprises the following steps:
step 31: extracting DNA of a related detection object;
step 32: performing sample quality detection on the extracted DNA;
step 33: designing and synthesizing a detection probe;
step 34: carrying out a preliminary experiment to obtain corresponding experiment basic data;
step 35: detecting by a fluorescent quantitative PCR instrument;
step 36: and (5) completing detection, and analyzing data to obtain a detection result.
The detection method for the MassARRAY method comprises the following steps:
step 41: preparing a detection sample;
step 42: carrying out the integrity detection of the sample and the pretreatment of the sample;
step 43: designing and synthesizing a primer;
step 44: performing PCR amplification, purification, extension or transcriptional cleavage based on Step 43;
step 45: performing spotting and mass spectrometry on the detection sample finished in the Step 44;
step 46: and obtaining a detection result.
The detection method for detecting the Illumina BeadXpress method comprises the following steps:
step 51: customizing a gene chip;
step 52: preparing a gene sample, and performing quality detection on the prepared sample;
step 53: carrying out on-machine detection on the sample by using a detection instrument;
step 54: analyzing the obtained detection data;
step 55: and obtaining a detection result.
2. The method of claim 1, wherein the in vitro protein binding assay is a method for detecting protein-protein interaction under in vitro conditions, and the target protein is obtained by SDS-page electrophoretic analysis, MS detection, protein-protein interaction verification or screening.
3. The method of claim 1, wherein the in vitro protein binding assay is an in vitro validation/search for interacting proteins using a labeled antibody.
4. The method of claim 1, wherein the protein is combined with a biotin-labeled RNA/DNA probe in vitro, and incubated with the cytoplasmic protein extract to form an RNA/DNA-protein complex.
5. The method of claim 1, wherein the co-immunoprecipitation is a method for studying protein interactions based on the interaction between an antibody and an antigen.
6. The method for detecting biological molecules as claimed in claim 1, wherein the probe method detection is based on fluorescent quantitative PCR, and PCR primers and PCR amplifications are designed for different SNP sites on the chromosome respectively.
7. The method of claim 1, wherein the MassARRAY method comprises exciting DNA molecule fragments with laser, and determining the molecular weight of the fragments by flight time.
8. The method of claim 1, wherein the Illumina BeadXpress method is suitable for high throughput detection, and can simultaneously detect a plurality of SNP sites for genomic chip result confirmation.
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