CN104313124A - Specific primer for detecting bovine IGF-1 genes, fluorescent quantitative PCR detection kit of bovine GHR genes, and detection method and application of kit - Google Patents
Specific primer for detecting bovine IGF-1 genes, fluorescent quantitative PCR detection kit of bovine GHR genes, and detection method and application of kit Download PDFInfo
- Publication number
- CN104313124A CN104313124A CN201410253252.5A CN201410253252A CN104313124A CN 104313124 A CN104313124 A CN 104313124A CN 201410253252 A CN201410253252 A CN 201410253252A CN 104313124 A CN104313124 A CN 104313124A
- Authority
- CN
- China
- Prior art keywords
- igf
- genes
- kit
- gene
- bovine
- 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
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/6851—Quantitative amplification
Abstract
The invention relates to a specific primer for detecting bovine IGF-1 genes, a fluorescent quantitative PCR detection kit of the bovine GHR genes, and a detection method and an application of the kit. The fluorescent quantitative PCR detection kit of the bovine IGF-1 genes comprises a PCR reaction solution, a TaqDNA polymerase mixture, a SYBRGreen dye and primers. The sequences of the primers are as follows: the sequence of an upstream primer is 5'-AGCAGTCTTCCAACCCAA-3 '; and the sequence of a downstream primer is 5'-AGATGCGAGGAGGATGTG-3'. The invention also provides a detection method and an application of the above kit and the specific primer for detecting the bovine IGF-1 genes. The object of conveniently and rapidly detecting an mRNA expression level of the genes is achieved. The detection kit has the advantages of high sensitivity, good stability and low experimental cost in detecting the mRNA expression level of the genes. The method provided by the invention can effectively amplify IGF-1 genes derived from three bovine species including dairy cow, cattle and yak, and can detect the mRNA expression quantity of the IGF-1 genes.
Description
Technical field
The invention belongs to gene engineering technology field, be specifically related to a kind of Auele Specific Primer and the ox IGF-1 gene by fluorescence quantitative PCR detection kit that detect ox IGF-1 gene.
Background technology
(Polymerase Chain Reaction, PCR) is technology the most frequently used in DNA manipulation in vitro technology in polymerase chain reaction.Template DNA, Auele Specific Primer, dNTPs substrate, hot resistant DNA polymerase, magnesium ion etc. are placed in same buffering reaction system by round pcr, carry out repeatedly high-temperature denatured, low-temperature annealing, middle temperature chain extension thermal cycling, realize target dna fragment in reaction solution in 2
ndoubly amplification (wherein n is times of thermal cycle).
Fluorescent quantitative PCR technique is on the basis that flexible PCR reacts, in conjunction with real-time fluorescence detection technique and Computer Analysis technology.Quantitative fluorescent PCR adds specific fluorescent mark material in flexible PCR reaction system, the amplification situation of real time monitoring of DNA is carried out by the changing conditions of the fluorescent value in PCR reaction system after detecting each thermal cycling, and obtain the fluorescent quantitation change curve of each sample, and then obtain the Ct value (change in fluorescence reaches cycle number during threshold value) of each reaction tubes; Simultaneously, the target DNA of the dose known amounts of 2-10 multiple dilutions is detected under identical reaction system and reaction conditions, obtain its Ct value, with the logarithm of starting template number for X-coordinate, with Ct value for ordinate zou preparation standard curve, the initiate dna template number of Ct value to each reaction of typical curve and each sample carries out quantitative assay accordingly.
Although the mrna expression level of Northern blot technology detectable gene, whole experimentation operation more complicated.During Northern blot tests, a main problem is the degraded that there is RNA, so experimental articles all in Northern Blot all needs the process through removing RNA enzyme, as high bake, DEPC process etc.In addition, in Northern Blot, a lot of experimental article such as formaldehyde, EB, DEPC, ultraviolet lamp etc. have certain injury to human body.Although gene chip experiment reduces the workload of experimenter, and in once testing, can detect several thousand gene expression amounts changes, its sensitivity is lower simultaneously.
And SYBR Green is a kind of combination dye be incorporated in double-stranded DNA ditch.After being combined with double-stranded DNA, its fluorescence strengthens greatly.The detection that this character is used for amplified production is ideal.The maximum absorption wavelength of SYBR Green is about 497 nm, and emission wavelength is maximum is about 520 nm.In PCR reaction system, add excessive SYBR fluorescence dye, after SYBR fluorescence dye mixes DNA double-strand specifically, emitting fluorescence signal, and the SYBR dye molecule do not mixed in chain can not launch any fluorescent signal, thus ensure the increase of fluorescent signal and the increase Complete Synchronization of PCR primer.
SYBR Green has many good qualities in the real-time context of detection of nucleic acid, and because it combines with all double-stranded DNAs, need not customize especially because template is different, the program versatility therefore designed is good, and price is relatively low.In addition, because a PCR primer can be combined with polymolecular dyestuff, therefore the sensitivity of SRYB Green is very high.But because SYRB Green combines with all double-stranded DNAs, the false positive therefore caused by the amplified production of primer dimer, strand secondary structure and mistake can affect quantitative accuracy.The impact reducing non-specific product can be helped by the change of fluorescence after measurement raised temperature.The homogeneity carrying out assay products by melting curve contributes to analyzing and obtains quantitative result by SYBR Green.
Para-insulin growth factor (Insulin-Like Growth Factor-1, IGF-1) is the important excreted factor regulating animal growth.IGF-1 shows important regulating effect in embryo and postnatal growth course, and it can by running balance and the function of organization regulating the propagation of cell, differentiation and apoptosis to affect various tissue.Therefore, in the feeding process of ox and in scientific research, can the expression amount of IGF-1 in detection by quantitative tissue, reach the object of the development condition of monitoring ox.
Summary of the invention
The invention provides the fluorescent quantificationally PCR detecting kit of a kind of para-insulin growth factor (Insulin-Like Growth Factor-1, IGF-1) gene.By a large amount of experimental studies, our optimization is a set of effectively can detect the primer of different ox kind (comprising milk cow, ox and yak) IGF-1 gene expression amount and the condition of PCR reaction, and be assembled into IGF-1 genetic expression and detect fluorescent quantitative poly chain reaction test kit, to meet the detection needs of life science, medical science and animal and veterinary investigator.For this reason, the present invention also provides the application of Auele Specific Primer, detection method and the test kit detecting ox IGF-1 gene.
In order to realize foregoing invention object, technical scheme provided by the invention is as follows:
Detect an Auele Specific Primer for ox IGF-1 gene, its nucleotides sequence is classified as:
Upstream primer sequence is 5 '-AGCAGTCTTCCAACCCAA-3 ';
Downstream primer sequence is: 5 '-AGATGCGAGGAGGATGTG-3 '.
Second object of the present invention is to provide a kind of fluorescent quantificationally PCR detecting kit of ox IGF-1 gene, and described test kit comprises PCR reaction solution, Taq DNA polymerase mixture, SYBR Green dyestuff and primer; Described primer sequence is:
Upstream primer sequence is 5 '-AGCAGTCTTCCAACCCAA-3 ';
Downstream primer sequence is: 5 '-AGATGCGAGGAGGATGTG-3 '.
Preferably, described test kit also comprises positive control and negative control.
3rd object of the present invention is to provide the quantitative detecting method of ox IGF-1 gene, for template with cDNA to be measured, the primer described in claim 1 and SYBR Green dyestuff is utilized to carry out real-time fluorescence quantitative PCR, record Ct value, and using IGF-1 standard gene as positive control drawing standard curve, typical curve and Ct value carry out quantitative assay accordingly.
Preferably, the reaction conditions of described real-time fluorescence quantitative PCR is: 95 DEG C of denaturations 30 seconds; 95 DEG C 5 seconds, 56 DEG C 20 seconds, catch fluorescent signal, circulate 40 times.
4th object of the present invention is to provide the application of mentioned reagent box in detection by quantitative ox IGF-1 gene.
Preferably, described ox is milk cow, ox and yak.
Compared with existing best technique, the invention has the advantages that:
1. present invention achieves the object conveniently detecting IGF-1 mrna expression.
2. the present invention has highly sensitive, that good stability, experimental cost are low advantage in detection mrna expression.
3. due to primer complete homology in milk cow, ox, yak, effectively can increase with the inventive method and derive from the IGF-1 gene of these species, thus detect the expression amount of IGF-1 gene.
Accompanying drawing explanation
Accompanying drawing is used to provide a further understanding of the present invention, and forms a part for specification sheets, together with embodiments of the present invention for explaining the present invention, is not construed as limiting the invention.In the accompanying drawings:
Fig. 1 is the fluoroscopic examination result figure of IGF-1 gene transcription level, A: amplification curve.X-coordinate is PCR cycle index, and ordinate zou is relative fluorescence value (Relative Fluorescence Units, RFU), B: melting curve, and X-coordinate is temperature, and ordinate zou is the variable quantity of relative fluorescence at unit temperature.
Embodiment
Following embodiment is convenient to understand the present invention better, but does not limit the present invention.Experimental technique in following embodiment, if no special instructions, is ordinary method.Test materials used in following embodiment, if no special instructions, is and purchases available from routine biochemistry reagent shop.
The fluorescent quantificationally PCR detecting kit of IGF-1 gene of the present invention is composed of the following components:
2×SYBR GREEN MIX 5mL；
Primer mixed solution 500 μ L;
Ultrapure water 5mL;
Positive control: standard I GF-1 gene template 500 μ L.
Wherein, 2 × SYBR GREEN MIX is composed of the following components: Taq archaeal dna polymerase 0.1 U/ μ L, dNTPs substrate 0.4 mmol/L, Mg
2+5.0 mmol/L, 100 mmol/L KCl, 20 mmol/L Tris. HCl PH8.3,0.02 % gelatin and 20 ng/mL SYBR Green dyestuffs.
Primer mixed solution: upstream primer 8 μm of ol/L, downstream primer 8 μm of ol/L, upstream primer sequence is 5 '-AGCAGTCTTCCAACCCAA-3 ', and downstream primer sequence is: 5 '-AGATGCGAGGAGGATGTG-3 '.
Standard I GF-1 gene template: concentration is 0.8 μm of ol/L, IGF-1 gene template sequence is 5 '-AGCAGTCTTCCAACCCAATTATTTAAGTGCTGCTTTTGTGATTTCTTGAAGCAGGT GAAGATGCCCATCACATCCTCCTCGCATCT-3 '.
Ultrapure water: purity is more than 18.25 M Ω .CM.
The step applying fluorescent quantificationally PCR detecting kit detection by quantitative ox IGF-1 gene of the present invention is as follows:
(1) get 2 × SYBR GREEN MIX 1000 μ L, primer mixed solution 100 μ L, ultrapure water 800 μ L fully mixes, the FQ-PCR preparing 1900 μ L reacts premixed liquid;
(2) fully mix above various liquid, premixed liquid is distributed into 100 tubules by 19 μ L/ pipes, adds in the special PCR reaction tubes of fluorescent quantitation or Sptting plate;
(3) preparation is managed with the standard I GF-1 gene template 5 of the dilution series such as 10E+8/ML, 10E+7/ML, 10E+6/ML, 10E+5/ML, 10E+4/ML, often pipe 10 μ L;
(4) 1 μ L cDNA to be measured, ultrapure water (negative control) or standard I GF-1 gene template is added respectively each being equipped with in the reaction tubes of 19 μ L FQ-PCR reaction premixed liquids, concussion mixing, after brief centrifugation, upper machine is in quantitative real time PCR Instrument (Bio-Rad CFX96TM), 95 DEG C of denaturations 30 seconds; 95 DEG C 5 seconds, 56 DEG C 20 seconds, catch fluorescent signal, circulate 40 times; Because the fragment of amplification is shorter, annealing and extension merge into a step, namely 56 DEG C, 20 seconds; Be increased to 95 DEG C, in temperature-rise period, catch fluorescent signal, make solubility curve according to this fluorescent signal.Pcr amplification curve shows IGF-1 gene by fluorescence signal value standard compliant " S " type curve see Fig. 1, Fig. 1, and melting curve shows that this fluorescent quantitation has the detection specificity of height.Therefore, the present invention can measure the expression level of IGF-1 accurately, and has the specificity of height, and effect is very desirable, completely the same with the expection of design.
Wherein, cDNA to be measured is prepared by following steps:
A, RNA leaching process: the tissue gathering ox kind (ox, milk cow or yak), is organized in 100mg in liquid nitrogen and grinds, add 1mL TRIzol, carry out homogenized with Syrup-homogenizing instrument; Homogenised sample is placed 5 minutes in room temperature (15-30 DEG C), nucleic acid-protein mixture is separated completely; Add 0.2mL chloroform, thermal agitation 15 seconds, room temperature places 3 minutes; Centrifugal 15 minutes (sample is divided into three layers: bottom is yellow organic phase, and upper strata is colourless aqueous phase and a middle layer) of 4 DEG C of 10000 × g; Aqueous phase is transferred in new pipe; Add the RNA in 0.5mL isopropanol precipitating aqueous phase, room temperature places 10 minutes; , after centrifugal, at the bottom of pipe side and pipe, there is gelatinous precipitate in centrifugal 10 minutes of 4 DEG C of 10000 × g; Remove supernatant, use 1mL 75% washing with alcohol RNA precipitation; Centrifugal 5 minutes of 4 DEG C of 7000 × g, abandon supernatant; Room temperature is placed dry RNA and is precipitated; Adding 25-200 μ l beats several times without the water rifle head suction of RNase, and 58 DEG C of placements make RNA dissolve in 10 minutes, now obtain the RNA solution of ox kind.
B, transcriptive process,reversed, namely obtain the process of cDNA: in PCR pipe, add 50 μm of ol/L oligo dT solution 1.5 μ L, 10 mmol/L dNTP mixed solution 1 μ L, yak total serum IgE 2.5 μ L, RNase free dH
2o 5 μ L, mixing, places 5min, is transferred to cooled on ice rapidly for 65 DEG C; Continue to add 5 × Prime ScriptTM damping fluid 4 μ L in above-mentioned PCR pipe, 40 U/ μ l RNA enzyme inhibitors 0.5 μ L, 200 U/ μ L Prime Script
tMthermoScript II 1 μ L, RNase free dH
2o 4.5 μ L, mixing, 30 DEG C of 10min, 42 DEG C of 60min, react under 70 DEG C of 15min conditions, now obtains the cDNA of ox kind.
(5) after PCR reaction terminates, read the Ct value of each reaction in order to quantitative analysis: with the logarithm of starting template number for X-coordinate, with the Ct value of positive control for ordinate zou preparation standard curve, the starting template number of Ct value to each reaction of typical curve and each sample carries out quantitative assay accordingly.Carry out PCR primer electrophoresis, ethidium bromide staining with 3% sepharose simultaneously, observe under gel imaging system, the result of each PCR reaction of gray scale scanning analytic record.To determine the size of amplified production, thus determine further increase whether be goal gene.
Due to primer complete homology in milk cow, ox, yak, effectively can increase with the inventive method and derive from the IGF-1 gene of these species, thus detect the expression amount of IGF-1 gene.
Last it is noted that the foregoing is only the preferred embodiments of the present invention, be not limited to the present invention, although with reference to previous embodiment to invention has been detailed description, for a person skilled in the art, it still can be modified to the technical scheme described in foregoing embodiments, or carries out equivalent replacement to wherein portion of techniques feature.Within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (7)
1. detect an Auele Specific Primer for ox IGF-1 gene, its nucleotides sequence is classified as:
Upstream primer sequence is 5 '-AGCAGTCTTCCAACCCAA-3 ';
Downstream primer sequence is: 5 '-AGATGCGAGGAGGATGTG-3 '.
2. a fluorescent quantificationally PCR detecting kit for ox IGF-1 gene, is characterized in that: described test kit comprises PCR reaction solution, Taq archaeal dna polymerase mixture, SYBR Green dyestuff and primer; Described primer sequence is:
Upstream primer sequence is 5 '-AGCAGTCTTCCAACCCAA-3 ';
Downstream primer sequence is: 5 '-AGATGCGAGGAGGATGTG-3 '.
3. test kit according to claim 2, is characterized in that: described test kit also comprises positive control and negative control.
4. the quantitative detecting method of an ox IGF-1 gene, for template with cDNA to be measured, the primer described in claim 1 and SYBR Green dyestuff is utilized to carry out real-time fluorescence quantitative PCR, record Ct value, and using IGF-1 standard gene as positive control drawing standard curve, typical curve and Ct value carry out quantitative assay accordingly.
5. detection method according to claim 4, is characterized in that: the reaction conditions of described real-time fluorescence quantitative PCR is: 95 DEG C of denaturations 30 seconds; 95 DEG C 5 seconds, 56 DEG C 20 seconds, catch fluorescent signal, circulate 40 times.
6. the application of the test kit described in Claims 2 or 3 in detection by quantitative ox IGF-1 gene.
7. application according to claim 6, is characterized in that: described ox is milk cow, ox and yak.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410253252.5A CN104313124A (en) | 2014-06-10 | 2014-06-10 | Specific primer for detecting bovine IGF-1 genes, fluorescent quantitative PCR detection kit of bovine GHR genes, and detection method and application of kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410253252.5A CN104313124A (en) | 2014-06-10 | 2014-06-10 | Specific primer for detecting bovine IGF-1 genes, fluorescent quantitative PCR detection kit of bovine GHR genes, and detection method and application of kit |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104313124A true CN104313124A (en) | 2015-01-28 |
Family
ID=52368433
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410253252.5A Pending CN104313124A (en) | 2014-06-10 | 2014-06-10 | Specific primer for detecting bovine IGF-1 genes, fluorescent quantitative PCR detection kit of bovine GHR genes, and detection method and application of kit |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104313124A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105039558A (en) * | 2015-08-11 | 2015-11-11 | 中国农业科学院兰州畜牧与兽药研究所 | Cattle HIF-1A gene transcriptional level fluorescent quantization PCR detection kit |
| CN105039557A (en) * | 2015-08-11 | 2015-11-11 | 中国农业科学院兰州畜牧与兽药研究所 | Cattle PPARG gene transcriptional level fluorescent quantization PCR detection kit |
| CN105039559A (en) * | 2015-08-11 | 2015-11-11 | 中国农业科学院兰州畜牧与兽药研究所 | Cattle FABP3 gene transcriptional level fluorescent quantization PCR detection kit |
| CN107586853A (en) * | 2017-09-12 | 2018-01-16 | 苏州博尔达生物科技有限公司 | Primer, probe, kit and its application based on RPA technology for detection calf-derived Cyclosporas |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101634657A (en) * | 2009-04-14 | 2010-01-27 | 李彬 | Preparation method of adipocytes differentiation metabolic product antibody chip |
| CN103275211A (en) * | 2013-06-17 | 2013-09-04 | 吉林大学 | Method for extracting IGF-1 (Insulin-like Growth Factor-1) from deer antler lyophilized powder |
-
2014
- 2014-06-10 CN CN201410253252.5A patent/CN104313124A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101634657A (en) * | 2009-04-14 | 2010-01-27 | 李彬 | Preparation method of adipocytes differentiation metabolic product antibody chip |
| CN103275211A (en) * | 2013-06-17 | 2013-09-04 | 吉林大学 | Method for extracting IGF-1 (Insulin-like Growth Factor-1) from deer antler lyophilized powder |
Non-Patent Citations (1)
| Title |
|---|
| JIE PEI等: "Expression profiles of growth hormone receptor and insulin-like growth factor I in cattle and yak tissues revealed by quantitative real-time PCR", 《ARCHIV TIERZUCHT》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105039558A (en) * | 2015-08-11 | 2015-11-11 | 中国农业科学院兰州畜牧与兽药研究所 | Cattle HIF-1A gene transcriptional level fluorescent quantization PCR detection kit |
| CN105039557A (en) * | 2015-08-11 | 2015-11-11 | 中国农业科学院兰州畜牧与兽药研究所 | Cattle PPARG gene transcriptional level fluorescent quantization PCR detection kit |
| CN105039559A (en) * | 2015-08-11 | 2015-11-11 | 中国农业科学院兰州畜牧与兽药研究所 | Cattle FABP3 gene transcriptional level fluorescent quantization PCR detection kit |
| CN105039558B (en) * | 2015-08-11 | 2018-06-15 | 中国农业科学院兰州畜牧与兽药研究所 | Ox HIF-1A gene transcription level fluorescent quantificationally PCR detecting kits |
| CN107586853A (en) * | 2017-09-12 | 2018-01-16 | 苏州博尔达生物科技有限公司 | Primer, probe, kit and its application based on RPA technology for detection calf-derived Cyclosporas |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US10683538B2 (en) | Quality assessment of circulating cell-free DNA using multiplexed droplet digital PCR | |
| CN103667514B (en) | A kind of human interleukin 2 8B gene pleiomorphism fluorescence PCR detection reagent kits | |
| CN104313124A (en) | Specific primer for detecting bovine IGF-1 genes, fluorescent quantitative PCR detection kit of bovine GHR genes, and detection method and application of kit | |
| CN105734158A (en) | Fluorescent PCR (polymerase chain reaction) detection kit for babesia caballi disease | |
| CN105586422A (en) | Kit for detecting TERT gene promoter mutation, and detection method and application of kit | |
| CN103103259A (en) | Method, kit and primers for determining whether two predetermined loci of nucleic acid sample mutate or not | |
| Murray et al. | A circulating microRNA panel for malignant germ cell tumor diagnosis and monitoring | |
| CN106967810A (en) | The method and kit of a kind of detection FGFR3 gene mutation diagnosing bladder cancers | |
| CN104894266B (en) | A kind of specific primer and fluorescence quantitative detection kit for detecting ox MSTN mrna expressions | |
| CN105950766A (en) | Primer group and kit for detecting HLA-B*5801 allelic genes | |
| CN104894120A (en) | Specific primers for detecting mRNA expression level of bovine LPL (lipoprotein lipase) genes and fluorescent quantitative detecting kit | |
| CN102676661B (en) | Fluorescent polarization based homogeneous phase detection method of single nucleotide polymorphism of codon118 of ERCC1 (excision repair cross-complementing 1) gene | |
| CN104278106B (en) | Duplex fluorescence quantitative RT-PCR (reverse transcription-polymerase chain reaction) detection kit for duck tembusu virus and egg drop syndrome virus | |
| CN103740822B (en) | Fluorescence quantitative PCR (polymerase chain reaction) combined rapid detection kit and method for SPF (specific pathogen free) mice pathogenic bacteria nucleic acid | |
| CN103725787B (en) | Fluorescent PCR (Polymerase Chain Reaction) detection kit for fusion genes of leukemia | |
| CN104313125A (en) | Specific primer for detecting bovine GHR genes, fluorescent quantitative PCR detection kit of bovine GHR genes, and detection method and application of kit | |
| CN103290134B (en) | Fluorogenic quantitative PCR detection kit for bovine ACTB gene transcription level | |
| KR20220034618A (en) | System for evaluating false positives due to contamination of positive control template using contamination index | |
| CN104894267A (en) | Specific primer for detecting mRNA expression levels of MYF5 genes of cows and fluorescent quantitative detecting kit | |
| CN104962629A (en) | Specific primer and fluorescent quantitative detection kit detecting cattle ADIPOQ gene mRNA expression level | |
| CN109593835B (en) | Method, kit and application for evaluating trace FFPE RNA sample | |
| JP2022517008A (en) | Methods and systems for identifying spinal muscular atrophy | |
| CN105039559A (en) | Cattle FABP3 gene transcriptional level fluorescent quantization PCR detection kit | |
| O’Neill et al. | Profiling circulating miRNAs from the plasma of individuals with metabolic syndrome | |
| CN103305608B (en) | Ox GAPDH gene transcription level fluorescent quantificationally PCR detecting kit |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20150128 |
|
| RJ01 | Rejection of invention patent application after publication |