CN108531548A - A kind of cell cDNA chip and its preparation method and application - Google Patents
A kind of cell cDNA chip and its preparation method and application Download PDFInfo
- Publication number
- CN108531548A CN108531548A CN201810311259.6A CN201810311259A CN108531548A CN 108531548 A CN108531548 A CN 108531548A CN 201810311259 A CN201810311259 A CN 201810311259A CN 108531548 A CN108531548 A CN 108531548A
- Authority
- CN
- China
- Prior art keywords
- cdna
- cell
- chip
- house
- primer
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- 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
- C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- 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
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/6851—Quantitative amplification
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Analytical Chemistry (AREA)
- Biophysics (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Physics & Mathematics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention discloses a kind of cell cDNA chip, including substrate and the cDNA templates that are fixed on substrate；The cDNA templates are the cDNA that the total serum IgE extracted by cell strain or primary cell passes through that reverse transcription obtains；It is a pair of the forward direction and reverse primer of house-keeping gene that the cDNA chip, which also has house-keeping gene primer, the house-keeping gene primer,.The cell cDNA chip of the present invention may be directly applied to cell strain screening, the Preparatory work of experiment time is short, first use one is only needed to open chip, save experiment reagent dosage, the difficulty of researcher's a large amount of cell culture and sample extraction is efficiently solved, the experiment condition of each sample is consistent, and experimental error is small, the accuracy and screening efficiency for improving born of the same parents' strain screening, can be used for the high flux screening of cell strain.The present invention also provides the preparation method and applications of the cell cDNA chip.
Description
Technical field
The present invention relates to genetic chip fields, more particularly, to a kind of cell cDNA chip and its preparation method and application.
Background technology
The life science of cell biology level and medical research are mainly from eucaryotic cell structure, cell function, albumen and each
Three aspect expansion of expression of the kind nucleic acid in cell.For comprehensively solve Scientific Research Problem, various hypothesis and supposition are explained, above three
A aspect complements each other, indispensable.From the point of view of experimental viewpoint, the expression for detecting various nucleic acid in cell is that cell biology is ground
Study carefully a most basic ring.Therefore, the primary according to being exactly of suitable cell strain or primary cell is selected：Target gene or albumen
Expression should meet experiment needs.
To select suitable cell strain or primary cell, researcher carries out the side of generally use when cell strain screening at present
Method is to buy or recover cell strain 2-5 plants to be selected, the culture up to one week to two weeks or more, amplification is first carried out, then to every plant
Different cells carries out RNA extractings, reverse transcription and target gene amplification.The above method has the following disadvantages：1) step is more, takes
It is long；2) screening efficiency is extremely low, can not real high flux screening；3) each cell strain or primary cell to be selected need to individually press above-mentioned side
Method is operated, and experiment condition is difficult to keep consistent, and the homogeneity between group is poor, influences the accuracy of the selection result；4) by not
With the cell strain especially limitation of primary cell sources and the limitation of experiment fees, researcher generally can not obtain simultaneously
All cell strains for obtaining targeted species, can only be screened in limited range of choice, and the cell category of detection is not complete, to
Influence subsequent experimental result.
Existing biochip technology is mostly that the DNA fragmentation (gene probe) of a large amount of known array is fixed on substrate
On, a two-dimentional DNA probe array is constituted, disposably a large amount of sequences in sample can be examined according to sequencing by hybridization method
It surveys and analyzes, can be used for gene expression profile measurement, abrupt climatic change, polymorphism analysis, genomic library mapping and sequencing by hybridization etc.,
But this method is not suitable for directly applying to the screening of various kinds of cell strain.
Therefore, the present situation screened for cell strain in the scientific research activity of this field, it is urgent to provide one kind to screen for cell strain
Novel research tool, be applicable to cell strain multisample analysis and quickly molecular level analysis.
Invention content
Technical problem to be solved by the present invention lies in provide a kind of cell cDNA chip for cell strain screening, energy
The efficiency and accuracy for enough improving cell strain screening, shorten screening time, can be used for the high flux screening of cell strain.
In order to solve the above technical problems, the present invention provides a kind of cell cDNA chip, including substrate and it is fixed on substrate
CDNA templates；
The cDNA templates are the cDNA that the total serum IgE extracted by cell strain or primary cell passes through that reverse transcription obtains；It is described
Cell strain or primary cell come from people/animal；
The cDNA chip also has a house-keeping gene primer, the house-keeping gene primer be a pair of house-keeping gene it is positive and
Reverse primer, the house-keeping gene primer is as internal reference and positive control.
Specifically, the point sample section with array distribution on the substrate, the point sample section is for fixing cDNA moulds
Plate；The cDNA templates are two or more, are derived from two or more different cell strain or primary cell；
The different cDNA templates in source are fixed on different point sample sections.The selection of cell strain or primary cell, can be according to client's need
Ask carry out personalized customization.
Specifically, the applied sample amount for the cDNA templates for being fixed on different point sample sections is consistent, and control internal reference qPCR knots
The fluctuation of fruit Ct values is no more than 2.
Specifically, the total serum IgE should control purity before for reverse transcription, limit ranging from：The ratio of OD260/OD280
The ratio that value is 2.0 ± 0.1, OD260/OD230 is 2.0-2.1.
Specifically, the total serum IgE should measure the integrality of RNA before for reverse transcription, it is more than 7 height to select RIN values
The RNA of quality carries out reverse transcription, obtains cDNA.
Specifically, the house-keeping gene is β-actin (β actins), positive and reverse primer is SEQ ID NO：1
The nucleotide chain of sequence shown in~2 or its complementary strand.Other house-keeping genes, such as GAPDH, U6 etc. also can be selected in the β-actin.
The sequence of the house-keeping gene β-actin (β actins) is SEQ ID NO：The nucleotide chain of sequence shown in 3 or its complementation
Chain.Wherein,
SEQ ID NO：1：GAAGAGCTAC GAGCTGCCTG A
SEQ ID NO：2：CAGACAGCAC TGTGTTGGCG
SEQ ID NO：3：GAAGAGCTAC GAGCTGCCTG ACGGCCAGGT CATCACCATT GG CAATGAGC
GGTTCCGCTG CCCTGAGGCA CTCTTCCAGC CTTCCTTCCT GGGCATGG AG TCCTGTGGCA TCCACGAAAC
TACCTTCAAC TCCATCATGA AGTGTGACGT GGA CATCCGC AAAGACCTGT ACGCCAACAC AGTGCTGTC
The present invention also provides a kind of preparation methods of cell cDNA chip, include the following steps：
1) acquisition of cDNA：It is extracted to obtain total serum IgE by people/cell lines or primary cell, be purified；Detection and screening
RNA, RNA purity limit ranging from：The ratio that the ratio of OD260/OD280 is 2.0 ± 0.1, OD260/OD230 is 2.0-2.1,
The RIN values of RNA integralities are more than 7；Reverse transcription is carried out using the RNA filtered out, obtains cDNA；
2) cDNA templates are prepared：The cDNA that step 1) obtains is quantified, with fixed applied sample amount point sample on substrate,
Ensure that the fluctuation of internal reference qPCR result Ct values is no more than 2；CDNA templates are prepared into after freeze-drying.
Specifically, 96 hole PCR plates, 384 hole PCR plates or other specification PCR plates can be selected in the substrate.The cell c
Corresponding PCR plate overlay film is covered in DNA chip.
The present invention also provides the methods for using cell cDNA chip to carry out cell strain screening, include the following steps：
1) PCR reactions are carried out：With the template that the fixed cDNA templates of cell cDNA chip are PCR reactions, with house-keeping gene
Primer is the primer of PCR reactions, configures PCR reaction system solution, is added into cDNA arrays, the cDN A templates of freeze-drying is made to fill
Divide dissolving；CDNA chip is put into PCR instrument progress PCR reactions, annealing temperature control when PCR cycle is less than positive and reversed
The Tm values (melting temperature of DNA) of primer；Upstream and downstream primer special can be combined with the destination region of cDNA during this, be gone forward side by side
Row nucleotide chain type polymerisation.
2) phosphor collection is carried out, by the method for real time fluorescent quantitative come the expression of testing goal gene.It calculates in each sample
The △ Ct values of target gene and house-keeping gene, can analyze expression of the target gene in different cell strains or primary cell,
To help to select suitable cell strain or primary cell.
Specifically, including archaeal dna polymerase in PCR reaction system solution, archaeal dna polymerase is free nucleic acid 5 prime excision enzyme activity, packet
Include 5 ' exonuclease activities and 3 ' exonuclease activities.
Cell cDNA chip provided by the invention may be directly applied to cell strain screening, need not move through cell strain culture,
Amplification, in the periods such as RNA extractings, reverse transcription, sample time is short, reduces scientific research time cost.
Cell cDNA chip provided by the invention only needs first use one to open slide when being screened for cell strain, and existing skill
The screening scheme of art is needed using more than ten even hundreds and thousands of slides, and present invention saves experimental implementation workloads, effectively
The screening time for shortening multisample amount, can help researcher disposably to be screened according to the demand to destination gene expression level
Various kinds of cell strain can be used for the high flux screening of cell strain.
Cell cDNA chip provided by the invention only needs first use portion reagent when being screened for cell strain, and existing skill
Being used when each cell strain screening to be selected of the screening scheme of art needs a reagent, and the present invention is greatly saved experiment reagent use
Amount saves scientific research cost.
Cell cDNA chip provided by the invention can be customized service according to the demand of client, efficiently solve research
The difficulty that personnel's large sample is collected.
Cell cDNA chip provided by the invention has used the cDNA templates of homogenization, and the R of cDNA is prepared for reverse transcription
NA also passes through strict quality control, and the ratio that the ratio of OD260/OD280 is 2.0 ± 0.1, OD260/OD230 is 2.0-2.1, R NA
The RIN values of integrality are more than 7, and experimental error is small, and experiment condition is consistent, and improve the accuracy of born of the same parents' strain screening.
Cell cDNA chip provided by the invention is capable of providing Detailed clinical and the follow-up of detailed cell strain or primary cell
Information, facilitates which kind of correlation analysis researcher carries out.
Cell cDNA chip provided by the invention be used for screen cell strain method, have detection flux it is big, sensibility is high,
It is easy to operate quickly, the advantages such as dosing accuracy is high, result reliability is strong.
Description of the drawings
Fig. 1 is the Ct values that qPCR detects house-keeping gene β-actin in the cell cDNA chip of 15 plants of lung adenocarcinoma cells
Block diagram.
Fig. 2 be in the cell cDNA chip of 15 plants of lung adenocarcinoma cells the Ct values of qPCR testing goal genes Fibulin5 with
The block diagram of the difference of the Ct values of house-keeping gene β-actin.
Specific implementation mode
Clear, complete description will be carried out to technical scheme of the present invention below, it is clear that described embodiment is this hair
Bright a part of the embodiment, instead of all the embodiments.Based on the embodiments of the present invention, those of ordinary skill in the art exist
The every other embodiment obtained under the premise of not making creative work, shall fall within the protection scope of the present invention.
The present invention cell cDNA chip include：Including substrate and the cDNA templates being fixed on substrate；
CDNA templates are that RNA (ribonucleic acid) reverse transcription extracted by mankind or animal cell strain or primary cell obtains
cDNA；
The cell cDNA chip of the present invention further includes house-keeping gene primer, and house-keeping gene β-actin (β actins) are just
To and reverse primer it is a pair of.
Wherein, the cDNA templates are to be taken out by cell strain or primary cell (or carrying out personalized customization according to customer demand)
The cDNA that the RNA reverse transcriptions carried are formed.
Wherein, the forward direction of the house-keeping gene β-actin (β actins) and reverse primer are SEQ ID NO：1-2 institutes
Show the nucleotide chain or its complementary strand of sequence.β-actin can also be replaced by other house-keeping genes, such as GAPDH, U6 etc..
Wherein, the sequence of the house-keeping gene β-actin (β actins) is SEQ ID NO：The nucleosides of sequence shown in 3
Sour chain or its complementary strand.
Sequence is described as follows：
SEQ ID NO：1：GAAGAGCTAC GAGCTGCCTG A
SEQ ID NO：2：CAGACAGCAC TGTGTTGGCG
SEQ ID NO：3：GAAGAGCTAC GAGCTGCCTG ACGGCCAGGT CATCACCATT GGCAATGAGC
GGTTCCGCTG CCCTGAGGCA CTCTTCCAGC CTTCCTTCCT GGGCATGGAG TCCTGTGGCA TCCACGAAAC
TACCTTCAAC TCCATCATGA AGTGTGACGT GGACATCCGC AAAGACCTGT ACGCCAACAC AGTGC TGTC
The cDNA templates of the cell cDNA chip of the present invention are to obtain by the following method：
1. people/cell lines or primary cell extract to obtain total serum IgE；
2.RNA is purified；
3.nanodrop carries out purity detecting, to select high-purity RNA, limits ranging from：260/280 for 2.0 (±
0.1), 260/230 is 2.0-2.1；
4. Agilent bioanalysis accurately measures the integrality of RNA, it is more than that the RNA of 7 high quality is carried out instead to select RIN values
Transcription, obtains cDNA；
5.cDNA is quantified by Qubit, and 96 orifice plate point samples are carried out with fixed applied sample amount, to ensure internal reference qPCR knots
The fluctuation of fruit Ct values is no more than 2.
It is anti-using archaeal dna polymerase, dNTP and corresponding PCR when the cell cDNA chip of the present invention is screened for cell strain
Buffer solution is answered accordingly to be detected by PCR.Wherein, PCR (the Polymerase Chain used
Reaction, PCR) annealing temperature in response procedures when PCR cycle should be less than it is positive and reverse primer
Tm (melting temperature of DNA) value.In the case of effectively expanding, sample detection credible result, otherwise experiment needs to repeat.
Detection method includes the following steps for cell cDNA chip of the present invention：
Include to be extracted by cell strain or primary cell in PCR system using PCR system
CDNA (cDNA of all samples is by accurate quantitative and homogenization), and the sense primer of a pair of specific house-keeping gene and
Downstream primer.
The basic step of reaction is as follows：First, cDNA chip is taken out from -20 DEG C, room temperature 1min is placed in, in centrifuge tube
Middle configuration reaction system solution；Then the sealer for removing cDNA chip is added after above-mentioned mixed liquor mixing with 20 holes μ l/
In cDNA arrays；CDNA chip is sealed with new sealer, pays attention to having to carefully being adjacent to, after sealing, orifice plate is positioned over ice
Upper 15min so that the cDNA of freeze-drying fully dissolves, and after gently shaking, 2000-6000rpm centrifuges 1min；Orifice plate is put into PCR
Instrument, sets program, carries out PCR experiment, and DNA double chain is opened by denaturation, according to the Tm values of primer select suitable annealing and
Elongating temperature carries out the annealing and extension of DNA, and upstream and downstream primer special can be combined with the destination region of cDNA during this, and
Nucleotide chain type polymerisation is carried out, phosphor collection is finally carried out；As a result, by the method for real time fluorescent quantitative come testing goal
The expression of gene；Finally, the differential expression of target gene is obtained by calculating target gene and the method for house-keeping gene △ Ct values
Situation.
An example to detect the differential expression of cancer related gene Fibulin5 illustrates：First, according to we
Method feature, the primer of design synthesis Fibulin5.By the preparation method of the present invention by obtaining cDNA in 15 kinds of lung adenocarcinoma cell lines
Template is simultaneously prepared into cDNA chip.Using house-keeping gene β-actin as internal reference, primer is with reference to SEQ ID NO：1-2 sequent synthesis.Expand
It is as follows to increase program：(1) PCR amplification system：PCR reaction solution is configured in accordance with the following methods.(2) PCR amplification program is as follows：According to
Lower method carries out PCR amplification, and PCR reaction conditions are：95 DEG C of pre-degenerations 10 minutes, 95 DEG C 15 seconds, 58 DEG C 40 seconds, amplified reaction 40
Cycle collects fluorescence 60 DEG C of 40 second stages.
Interpretation of result is as follows：Referring to Figures 1 and 2, according to △ Ct values [(cancer sample Ct values-internal reference Ct values)] icp gene
Expressions of the Fibulin5 in different cell strains.As shown in Figure 1,15 kinds of lung adenocarcinoma cell line warps in the cell cDNA chip
QPCR detects the Ct values fluctuation≤2 of house-keeping gene β-actin, illustrates the cell cDNA obtained by preparation method through the invention
The homogenization degree of chip is high, and experimental error is small between group, and the accuracy for screening cell strain is high.Can intuitively it be found out very much by Fig. 2,
It deducts after internal reference the expression of gene Fibulin5 in lung adenocarcinoma cell line D6 and is apparently higher than other cell strains, this method can be with
Disposable screening various kinds of cell strain, can be used for the high flux screening of cell strain.
In conclusion the various embodiments described above are only presently preferred embodiments of the present invention, not limiting the present invention's
Protection domain, all within the spirits and principles of the present invention, any modification, equivalent substitution, improvement and etc. done should be all included in
In protection scope of the present invention.
Sequence table
<110>Xinchao Biotech Co., Ltd., Shanghai
<120>A kind of cell cDNA chip and its preparation method and application
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 21
<212> DNA
<213>Artificial sequence (artificial sequence)
<400> 1
gaagagctac gagctgcctg a 21
<210> 2
<211> 20
<212> DNA
<213>Artificial sequence (artificial sequence)
<400> 2
cagacagcac tgtgttggcg 20
<210> 3
<211> 189
<212> DNA
<213>Artificial sequence (artificial sequence)
<400> 3
gaagagctac gagctgcctg acggccaggt catcaccatt ggcaatgagc ggttccgctg 60
ccctgaggca ctcttccagc cttccttcct gggcatggag tcctgtggca tccacgaaac 120
taccttcaac tccatcatga agtgtgacgt ggacatccgc aaagacctgt acgccaacac 180
agtgctgtc 189
Claims (10)
1. a kind of cell cDNA chip, which is characterized in that including substrate and the cDNA templates being fixed on substrate；
The cDNA templates are the cDNA that the total serum IgE extracted by cell strain or primary cell passes through that reverse transcription obtains；
It is that a pair of house-keeping gene is positive and reversed that the cDNA chip, which also has house-keeping gene primer, the house-keeping gene primer,
Primer, the house-keeping gene primer is as internal reference and positive control.
2. cell cDNA chip as described in claim 1, which is characterized in that the point sample with array distribution on the substrate
Section, the point sample section is for fixing cDNA templates；The cDNA templates be two or more, be derived from two kinds or
Two or more different cell strains or primary cell；The different cDNA templates in source are fixed on different point sample sections.
3. cell cDNA chip as described in claim 1, which is characterized in that be fixed on the cDNA templates in different point sample sections
Applied sample amount is consistent, and is controlled the fluctuation of internal reference qPCR result Ct values and be no more than 2.
4. cell cDNA chip as described in claim 1, which is characterized in that the total serum IgE should be controlled before for reverse transcription
Purity processed limits ranging from：The ratio that the ratio of OD260/OD280 is 2.0 ± 0.1, OD260/OD230 is 2.0-2.1.
5. cell cDNA chip as described in claim 1, which is characterized in that the total serum IgE should be surveyed before for reverse transcription
Determine the integrality of RNA, it is more than that the RNA of 7 high quality carries out reverse transcription to select RIN values, obtains cDNA.
6. cell cDNA chip as described in claim 1, which is characterized in that the house-keeping gene is β-actin, it is positive and
Reverse primer is SEQ ID NO：The nucleotide chain of sequence shown in 1~2 or its complementary strand.
7. a kind of preparation method of cell cDNA chip, which is characterized in that include the following steps：
1) acquisition of cDNA：It is extracted to obtain total serum IgE by cell strain or primary cell, be purified；Detection and screening RNA, RNA purity limit
Determine ranging from：The ratio that the ratio of OD260/OD280 is 2.0 ± 0.1, OD260/OD230 is 2.0-2.1, RNA integralities
RIN values are more than 7；Reverse transcription is carried out using the RNA filtered out, obtains cDNA；
2) cDNA templates are prepared：The cDNA that step 1) obtains is quantified, with fixed applied sample amount point sample on substrate, it is ensured that
The fluctuation of internal reference qPCR result Ct values is no more than 2；CDNA templates are prepared into after freeze-drying.
8. preparation method as claimed in claim 7, which is characterized in that the substrate selects 96 hole PCR plates or 384 hole PCR plates；
Corresponding PCR plate overlay film is covered on the cell cDNA chip.
9. the cell cDNA chip that a kind of preparation method by described in claim 7 is prepared.
10. a kind of method that cell cDNA chip using as described in claim 1 or 9 carries out cell strain screening, feature exist
In including the following steps：
1) PCR reactions are carried out：With the template that the fixed cDNA templates of cell cDNA chip are PCR reactions, with house-keeping gene primer
For the primer of PCR reactions, PCR reaction system solution is configured, is added into cDNA arrays, keeps the cDNA templates of freeze-drying fully molten
Solution；CDNA chip is put into PCR instrument progress PCR reactions, annealing temperature control when PCR cycle is less than positive and reverse primer
Tm values；Upstream and downstream primer special can be combined with the destination region of cDNA during this, and it is anti-to carry out the polymerization of nucleotide chain formula
It answers；
2) it carries out phosphor collection and calculates mesh in each sample by the method for real time fluorescent quantitative come the expression of testing goal gene
Mark the △ Ct values of gene and house-keeping gene.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810311259.6A CN108531548A (en) | 2018-04-09 | 2018-04-09 | A kind of cell cDNA chip and its preparation method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810311259.6A CN108531548A (en) | 2018-04-09 | 2018-04-09 | A kind of cell cDNA chip and its preparation method and application |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108531548A true CN108531548A (en) | 2018-09-14 |
Family
ID=63482325
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810311259.6A Pending CN108531548A (en) | 2018-04-09 | 2018-04-09 | A kind of cell cDNA chip and its preparation method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108531548A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005116261A2 (en) * | 2004-05-24 | 2005-12-08 | Albert Einstein College Of Medecine Of Yeshiva University | Rna expression microarrays |
| CN1952172A (en) * | 2005-10-20 | 2007-04-25 | 北京市农林科学院 | cDNA chip for determining gene expression information of crucifer under threatening conduction |
| CN103555839A (en) * | 2013-11-01 | 2014-02-05 | 东南大学 | Urinary sediment cell kidney fibrosis detection chip based on real-time fluorescent PCR (photo conductive relay) |
| CN103849679A (en) * | 2012-11-28 | 2014-06-11 | 康隽生物科技股份有限公司 | Detection method of enzyme type gene chip |
| CN106755330A (en) * | 2016-11-29 | 2017-05-31 | 上海芯超生物科技有限公司 | Cancer related gene differential expression detection kit and its application |
-
2018
- 2018-04-09 CN CN201810311259.6A patent/CN108531548A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005116261A2 (en) * | 2004-05-24 | 2005-12-08 | Albert Einstein College Of Medecine Of Yeshiva University | Rna expression microarrays |
| CN1952172A (en) * | 2005-10-20 | 2007-04-25 | 北京市农林科学院 | cDNA chip for determining gene expression information of crucifer under threatening conduction |
| CN103849679A (en) * | 2012-11-28 | 2014-06-11 | 康隽生物科技股份有限公司 | Detection method of enzyme type gene chip |
| CN103555839A (en) * | 2013-11-01 | 2014-02-05 | 东南大学 | Urinary sediment cell kidney fibrosis detection chip based on real-time fluorescent PCR (photo conductive relay) |
| CN106755330A (en) * | 2016-11-29 | 2017-05-31 | 上海芯超生物科技有限公司 | Cancer related gene differential expression detection kit and its application |
Non-Patent Citations (5)
| Title |
|---|
| AKIHIKO SAITO-HISAMINATO ET AL.: "Genome-Wide Profiling of Gene Expression in 29 Normal Human Tissues with a cDNA Microarray", 《DNA RESEARCH》 * |
| 张嵘: "《生物化学与分子生物学实验》", 31 August 2014, 中国医药科技出版社 * |
| 纪兆华等: "《差异表达基因检测统计方法研究》", 31 March 2017, 北京理工大学出版社 * |
| 范保星 等: "α粒子诱发BEP2D细胞转化过程中肺癌相关基因表达的cDNA microarray研究", 《CHINESE JOURNAL OF CANCER》 * |
| 袁媛 等: "《中药资源转录组分析操作指南》", 31 May 2016 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20220282303A1 (en) | Methods for standardized sequencing of nucleic acids and uses thereof | |
| CA2497988C (en) | Quantification of gene expression | |
| CN103898199B (en) | A kind of high-throughput nucleic acid analysis method and application thereof | |
| JP2018110597A (en) | Multivariate diagnostic assays and methods for using the same | |
| US20060286558A1 (en) | Normalization of samples for amplification reactions | |
| Cheesman et al. | Real-time quantitative PCR for analysis of genetically mixed infections of malaria parasites: technique validation and applications | |
| CN105899680A (en) | Nucleic acid probe and method of detecting genomic fragments | |
| CN107058538B (en) | Primer composition, kit composed of primer composition and application of kit | |
| US20220154261A1 (en) | Nucleic acid detection method and assay kit | |
| CN102634587B (en) | Method for combined and extended detection of continuous mutation of base by deoxyribonucleic acid (DNA) chips | |
| CN102337338A (en) | Method for simultaneously and quickly detecting number of five types of chromosomes, and kit and application thereof | |
| US7049104B2 (en) | Genetic analysis method | |
| CN108265113A (en) | ALDH2 genetic polymorphism detection kits | |
| Netto et al. | Diagnostic molecular pathology: current techniques and clinical applications, part I | |
| CN101805790A (en) | Method for simultaneously detecting polymorphism of 32 SNP loci on 24 sports-related genes | |
| CN108136389A (en) | Sample is to the automatic preparation in NGS libraries | |
| CN108384843A (en) | A method of identifying single copy gene mutated-genotype using digital pcr | |
| CN105543370B (en) | New comprehensive deaf-related gene mutation detection architecture and kit | |
| CN116445608B (en) | Composition for detecting deafness-related gene mutation, kit and application | |
| CN111394434A (en) | CHO host cell DNA residue detection kit of TaqMan probe method and application thereof | |
| CN108531548A (en) | A kind of cell cDNA chip and its preparation method and application | |
| Majewska et al. | Transcriptomic profiling during myogenesis | |
| CN106755330A (en) | Cancer related gene differential expression detection kit and its application | |
| US9528161B2 (en) | Materials and methods for quality-controlled two-color RT-QPCR diagnostic testing of formalin fixed embedded and/or fresh-frozen samples | |
| CN108642190B (en) | Forensic medicine composite detection kit based on 14 autosomal SNP genetic markers |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180914 |