CN108977514A - For screening the primer of wax gourd real-time fluorescence quantitative PCR reference gene EF-1 α - Google Patents
For screening the primer of wax gourd real-time fluorescence quantitative PCR reference gene EF-1 α Download PDFInfo
- Publication number
- CN108977514A CN108977514A CN201810919438.8A CN201810919438A CN108977514A CN 108977514 A CN108977514 A CN 108977514A CN 201810919438 A CN201810919438 A CN 201810919438A CN 108977514 A CN108977514 A CN 108977514A
- Authority
- CN
- China
- Prior art keywords
- primer
- wax gourd
- reference gene
- real
- gene
- 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.)
- Granted
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
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Biophysics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (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 invention belongs to technical field of molecular biology, and in particular to for screening the primer of wax gourd real-time fluorescence quantitative PCR EF-1 α reference gene.The present invention passes through the specific primer, screening obtains elongation factor-1α (Elongation Factor 1-alpha, EF-1 α) gene can stablize expression under wax gourd different tissues and low temperature stress, it is suitble in wax gourd gene expression research as reference gene.Elongation factor-1α (Elongation Factor 1-alpha, EF-1 α) gene is used for wax gourd gene expression analysis as reference gene for the first time by the present invention, is conducive to the stability, the reproducibility and reliability that improve the research of wax gourd gene expression analysis；Detection primer proposed by the present invention has specificity, greatly improves detection efficiency, improves the confidence level of testing result.
Description
Technical field
The invention belongs to field of biotechnology, and in particular to glimmering in real time under wax gourd different tissues and low temperature stress for screening
The primer of Fluorescent Quantitative PCR reference gene EF-1 α.
Background technique
Wax gourd (Benincasa hispida Cogn.) belong to the annual herbaceous plant of Curcurbitaceae Benincasa.Wax gourd
Flower, leaf, pericarp, pulp, seed are worth with good dietotherapeutic, can be used for treating the diseases such as cough, asthma.Meanwhile melon
In be rich in amino acid, vitamin C, phenols isoreactivity compound, these compounds have one tailor-made in terms of prevent chronic disease
With, and wax gourd is unique not fatty melon dish in melon dish, is rich in hydroxymalonic acid ingredient, glucide can be inhibited to be converted into rouge
Fat ingredient.With going deep into for research, wax gourd functional component and Regulation Mechanism related gene table are understood using Protocols in Molecular Biology
Reach for important research direction.
The research about the reference gene under the conditions of wax gourd different parts and low temperature stress has not been reported at present.The present invention
With respectively with " casting skin wax gourd " different tissues (root, stem, leaf, flower) and 4 DEG C of low-temperature treatments 0,1,6,12, lower 2 true leaves are for 24 hours
Material comments the expression stability analysis of 7 candidate reference genes using GeNorm, NormFinder and BestKeeper
Valence filters out elongation factor-1α (Elongation Factor 1-alpha, EF-1 α) gene as suitable internal reference base
Cause provides reference for the subsequent related gene expression research of wax gourd.
Summary of the invention
The object of the present invention is to provide for screening wax gourd different tissues real-time fluorescence quantitative PCR reference gene EF-1 α's
Primer, 1 pair of internal reference gene primer sequence specific amplification in the present invention is high, and amplification efficiency is close, and linear dependence is good, can use
The stable reference gene under screening wax gourd different tissues and low temperature stress.
The present invention is to solve above-mentioned technical problem by the following technical programs:
It is template, based on the sequencing of wax gourd transcript profile by wax gourd DNA, using Primer Premier5.0 software and follows
The principle of real-time fluorescence quantitative PCR design of primers designs 7 pairs of fluorescent quantitation special primers, is finally analyzed according to data, determines
This is the real-time fluorescence quantitative PCR primer to fluorescent quantitation special primer；
And the real-time fluorescence quantitative PCR primer are as follows:
Forward primer 5'- ACCGTCTATCAAGCACCC -3',
Reverse primer 5'- CCAAGTCCGCAGTCAAG -3'.
The beneficial effects of the present invention are:
The present invention is provided to screen the primer of wax gourd real-time fluorescence quantitative PCR reference gene, designed real time fluorescent quantitative
When PCR primer is used for wax gourd gene expression analysis, can be improved the stability of middle wax gourd gene expression analysis research, reliability and
Repeatability；In addition, designed real-time fluorescence quantitative PCR primer specificity is strong, so as to greatly improve using real-time
The detection efficiency of fluorogenic quantitative detection wax gourd, and improve the confidence level of testing result.
Detailed description of the invention
The invention will be further described in conjunction with the embodiments with reference to the accompanying drawings.
Fig. 1 is 7, wax gourd candidate reference gene amplifications.
Fig. 2A 28s candidate's reference gene qRT-PCR melt curve analysis.
Fig. 2 B EF-1 α candidate's reference gene qRT-PCR melt curve analysis.
Fig. 2 C GAPDH candidate's reference gene qRT-PCR melt curve analysis.
Fig. 2 D RP II candidate's reference gene qRT-PCR melt curve analysis.
Fig. 2 E TUA candidate's reference gene qRT-PCR melt curve analysis.
Fig. 2 F UBC21 candidate's reference gene qRT-PCR melt curve analysis.
Fig. 2 G UBQ candidate's reference gene qRT-PCR melt curve analysis.
Fig. 3 is 7 candidate reference gene mean CT-number distributions.
Fig. 4 GeNorm software analyzes the VN/VN+1 under different tissues and low temperature stress.
Specific embodiment
The present invention lists following representative embodiment, these embodiments are merely exemplary, and is not used in limitation
The scope of the present invention, these embodiments are only used for that the present invention will be described.And instrument employed in each embodiment and reagent
It is as follows: American AB I7500 real-time quantitative PCR instrument, American AB I Veriti 96-well thermal cycler, American AB I Power
SYBR Green PCR Master Mix, the first chain of cDNA synthetic agent box (PrimeScriptTM 1st Strand
CDNA Synthesis Kit), Marker DL 2000, Taq DNA Polymerase, dNTP be that precious bioengineering is (big
Even) Co., Ltd's product, primer synthesis are completed by Bo Shang biotechnology (Shanghai) Co., Ltd., remaining biochemical reagents is domestic
It analyzes pure.
The synthesis of embodiment 1 Total RNAs extraction and cDNA
Material to be tested " casting skin wax gourd " is uniformly seeded in the Fujian Academy crop town institute Fuqing City Dong Zhang vegetables scientific base.Cave
Disk nursery is respectively put into incubator when seedling grows to three leaves wholeheartedly and is handled.First group: normal cultivation condition, take root,
Stem, leaf spend four parts；Second group: 4 DEG C processing 0,1,6,12, for 24 hours, take 2 true leaves.Utilize BioTeke company generic plant
Total RNA extraction reagent box extracts RNA, and the agarose gel electrophoresis and UV1000 spectrophotometer for being 1% by concentration detect total
RNA integrality and concentration.Sample utilizes Takara company PrimeScript™ II 1st Strand cDNA Synthesis
The method of Kit kit synthesizes the first chain of cDNA, i.e., is cDNA by RNA reverse transcription；Later using gained cDNA as template, with reality
When fluorescence quantification PCR primer be that primer pair carries out PCR amplification, and the reaction system of PCR amplification and response procedures are as follows:
PCR reaction system: the total volume of reaction system is 25 μ L, positive and anti-containing 100 ng/ μ L cDNA, 2 μ L, 10 μm of ol/L
To each 1 μ L of primer, 10 mmol/L dNTP, 1 μ L, 5 U/0.2 μ L of μ L Taq archaeal dna polymerase, 1.5 mmol/L MgCl2
10 × PCR buffer, 2 μ L, remaining ingredient are the ultrapure water of sterilizing.
PCR response procedures are as follows: 94 DEG C of 3 min of initial denaturation；94 DEG C of 30 s of denaturation, 52 DEG C of 30 s of annealing, 72 DEG C extend 30
S, 35 circulations；Extend 7 min at last 72 DEG C；It is saved at 4 DEG C.
2 design of primers of embodiment and PCR primer verifying
Using seminar's early period to wax gourd transcript profile be sequenced gained gene order be template, filter out 28s, UBQ, RP II,
GAPDH, EF-1 α, UBC21, TUA 7 candidate reference genes utilize Primer Premier 5.0 according to design of primers principle
Design primer (table 1).Pcr amplification product is subjected to 1% agarose gel electrophoresis detection, testing result is as shown in Figure 1；It can by Fig. 1
Know, 7 reference gene amplified product bands are single, and size is identical as target fragment, illustrate primer energy specific amplification, are not present
Primer dimer is suitable for qRT-PCR and studies.
The candidate reference gene primer sequence of table 1
The verifying of 3 real-time fluorescence quantitative PCR primer of embodiment
Wax gourd total serum IgE is extracted, and according to the side of PrimeScriptTM 1st Strand cDNA Synthesis Kit kit
Method synthesizes the first chain of cDNA, i.e., is cDNA by RNA reverse transcription；Later using gained cDNA as template, according to Power SYBR
Green PCR Master Mix specification carries out PCR reaction on ABI7500 real-time quantitative PCR instrument, and PCR reacts anti-
Answer system as follows with response procedures:
Reaction system are as follows: the total volume of reaction system is 10 μ L, 5 μ L Power SYBR Green PCR Master
Mix, 2 μ L templates, the 0.4 μ L(concentration of forward primer of real-time fluorescence quantitative PCR primer is 10 μm of ol/ L in embodiment 2),
The 0.4 μ L(concentration of reverse primer of real-time fluorescence quantitative PCR primer is 10 μm of ol/ L in embodiment 2), distilled water is mended to totality
10 μ L of product.
Response procedures are as follows: 95 DEG C of initial denaturations 30s, PCR react 95 DEG C of 5s, 60 DEG C of 34s, 40 circulations.Melting curve journey
Sequence: 95 DEG C, 60 DEG C of 1min, 95 DEG C of 15s；4 DEG C of preservations；3 repetitions are done in each reaction.
For the result of reaction as shown in Fig. 2A-G, 7 candidate reference gene primer specificities are good, PCR melt curve analysis and only
There is a specific peak, shows primer free dimer, amplified band is single, high specificity, occur without non-specific amplification, thus
Show that seven pairs of designed primer specificities are strong (the real-time fluorescence quantitative PCR primer i.e. in embodiment 2), amplification efficiency is high, special
It is anisotropic strong, it can be used in the internal control primer experiment of wax gourd quantitative fluorescent PCR.
The analysis of 4 wax gourd reference gene expression stability of embodiment
The total serum IgE for extracting wax gourd root, stem, leaf, flower respectively, later respectively according to PrimeScriptTM 1st Strand cDNA
The method of Synthesis Kit kit synthesizes the first chain of cDNA, to obtain respective cDNA；Using real-time in embodiment 2
Fluorescent quantitation special primer is primer pair, using the cDNA of acquisition as template, carries out PCR amplification, PCR amplification system and expansion respectively
It is as follows to increase program:
PCR amplification system: the total volume of reaction system is 10 μ L, 5 μ L Power SYBR Green PCR Master
Mix, 2 μ L templates, the 0.4 μ L(concentration of forward primer of real-time fluorescence quantitative PCR primer is 10 μm of ol/ L in embodiment 2),
The 0.4 μ L(concentration of reverse primer of real-time fluorescence quantitative PCR primer is 10 μm of ol/ L in embodiment 2), distilled water is mended to totality
10 μ L of product.
PCR amplification program are as follows: 95 DEG C of 5 min of initial denaturation；95 DEG C of 5 s of denaturation, 60 DEG C of annealing 34s, 40 recycle；Last 72
Extend 10 min at DEG C；It is saved at 4 DEG C.
Using GeNorm, NormFinder and BestKeeper3 softwares to 7 candidate reference genes in different tissues
Expression stability assay, GeNorm software determine most suitable gene data, final GeNorm software analysis according to Fig. 4
As a result (table 2) it is found that in wax gourd different tissues 7 reference gene M values be ordered as UBQ=EF-1 α < RP II < UBC21 <
28s<TUA< GAPDH；NormFinder software analyzes (table 3), and candidate reference gene expression stationary value S is ordered as EF-1 α < RP
II<UBQ< UBC21<TUA<28s<GAPDH；BestKeeper software results (table 4) it is in the top be EF-1 α and RP
The SD value of II, TUA and GAPDH are small, but TUA correlation is poor, and GAPDH stability is worst, are negative value, are not suitable as internal reference
Gene.In summary known to three software analysis results: EF-1 α expression stability in different tissues is best.
Using GeNorm, NormFinder and BestKeeper3 softwares to 7 candidate reference genes in different tissues
Expression stability assay calculates 7 candidate reference gene expression stationary value (table 4), wax gourd low temperature using GeNorm software
Processing different periods M value is ordered as RP II=EF-1 α < UBC21 < UBQ < TUA < 28s < GAPDH, according to
NormFinder software is analyzed known to result (table 5), and 7 candidate reference gene expression are stablized under the conditions of 4 DEG C of low-temperature treatments
Value S is ordered as EF-1 α < RP II < TUA < 28s < UBC21 < UBQ < GAPDH;BestKeeper software directly utilizes CT
It is worth (Fig. 3) and carries out Data Management Analysis.The result shows that (table 6), EF-1 α is ranked first position under the conditions of low temperature stress, illustrates low
Temperature handles lower EF-1 α and is suitable as reference gene.In summary known to three software analysis results: EF-1 α is in low temperature stress
Expression stability is best.
2 GeNorm software of table analyzes 7 candidate reference gene expression stabilities under different tissues
Table 3NormFinder software analyzes 7 candidate reference gene expression stabilities under different tissues
4 BestKeeper software of table analyzes 7 candidate reference gene expression stabilities under different tissues
5 GeNorm software of table analyzes 7 candidate reference gene expression stabilities under the conditions of low temperature stress is tested
6 NormFinder software of table analyzes 7 candidate reference gene expression stabilities under the conditions of low temperature stress
7 BestKeeper software of table analyzes 7 candidate reference gene expression stabilities under the conditions of low temperature stress
To sum up, the present invention provides the real-time fluorescence quantitative PCR primer designed based on wax gourd reference gene, designed realities
When fluorescence quantification PCR primer be used for wax gourd gene expression analysis when, can be improved wax gourd gene expression analysis research stability,
Reproducibility and reliability；In addition, designed real-time fluorescence quantitative PCR primer specificity is strong, so as to greatly improve
Detection efficiency when wax gourd is detected using real time fluorescent quantitative, and improves the confidence level of testing result.
SEQUENCE LISTING
<110>Fujian Academy of Agricultural Sciences Crop Research Institute
<120>for screening the primer of wax gourd real-time fluorescence quantitative PCR reference gene EF-1 α
<130> 14
<160> 14
<170> PatentIn version 3.3
<210> 1
<211> 18
<212> DNA
<213>artificial sequence
<400> 1
aggaaggaga aggacgag 18
<210> 2
<211> 18
<212> DNA
<213>artificial sequence
<400> 2
ttcagaacca gacccaga 18
<210> 3
<211> 22
<212> DNA
<213>artificial sequence
<400> 3
gtgaattata gaatcgagca tc 22
<210> 4
<211> 20
<212> DNA
<213>artificial sequence
<400> 4
aaatcagaaa caatcccaac 20
<210> 5
<211> 19
<212> DNA
<213>artificial sequence
<400> 5
tcctccacaa tcgtctttc 19
<210> 6
<211> 17
<212> DNA
<213>artificial sequence
<400> 6
cccatcaccg ccataac 17
<210> 7
<211> 23
<212> DNA
<213>artificial sequence
<400> 7
tgtgaaactt gtctcgtggt atg 23
<210> 8
<211> 24
<212> DNA
<213>artificial sequence
<400> 8
tctcatcatc aggtccatct tact 24
<210> 9
<211> 18
<212> DNA
<213>artificial sequence
<400> 9
accgtctatc aagcaccc 18
<210> 10
<211> 17
<212> DNA
<213>artificial sequence
<400> 10
ccaagtccgc agtcaag 17
<210> 11
<211> 17
<212> DNA
<213>artificial sequence
<400> 11
attgggcgag gcgtatt 17
<210> 12
<211> 22
<212> DNA
<213>artificial sequence
<400> 12
ccctttgcac cttgacactt at 22
<210> 13
<211> 17
<212> DNA
<213>artificial sequence
<400> 13
gcaccgacct cctcata 17
<210> 14
<211> 19
<212> DNA
<213>artificial sequence
<400> 14
ccagccacct actgttgta 19
Claims (1)
1. the primer for screening wax gourd real-time fluorescence quantitative PCR reference gene EF-1 α, it is characterised in that: using wax gourd DNA as mould
Plate using Primer Premier5.0 software and follows real-time fluorescence quantitative PCR primer based on transcript profile result
The principle of design designs a pair of of fluorescent quantitation special primer, which is to screen wax gourd real-time fluorescence quantitative PCR EF-1 α
The primer of reference gene；Sequence is as follows:
Forward primer 5'- ACCGTCTATCAAGCACCC -3',
Reverse primer 5'- CCAAGTCCGCAGTCAAG -3'.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810919438.8A CN108977514B (en) | 2018-08-14 | 2018-08-14 | Primer for screening wax gourd real-time fluorescent quantitative PCR (polymerase chain reaction) reference gene EF-1 alpha |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810919438.8A CN108977514B (en) | 2018-08-14 | 2018-08-14 | Primer for screening wax gourd real-time fluorescent quantitative PCR (polymerase chain reaction) reference gene EF-1 alpha |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108977514A true CN108977514A (en) | 2018-12-11 |
| CN108977514B CN108977514B (en) | 2021-06-08 |
Family
ID=64552481
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810919438.8A Active CN108977514B (en) | 2018-08-14 | 2018-08-14 | Primer for screening wax gourd real-time fluorescent quantitative PCR (polymerase chain reaction) reference gene EF-1 alpha |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108977514B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110016518A (en) * | 2019-04-28 | 2019-07-16 | 福建省农业科学院作物研究所 | A kind of okra reference gene EF-1 α and its application |
| CN111575401A (en) * | 2020-07-07 | 2020-08-25 | 福建省农业科学院作物研究所 | Primer of towel gourd reference gene UBQ and application |
| CN111676230A (en) * | 2020-07-07 | 2020-09-18 | 福建省农业科学院作物研究所 | Towel gourd reference gene EF-1 alpha, and primer and application thereof |
-
2018
- 2018-08-14 CN CN201810919438.8A patent/CN108977514B/en active Active
Non-Patent Citations (5)
| Title |
|---|
| BIAO JIANG等: "De Novo Assembly and Characterization of the Transcriptome, and Development of SSR Markers in Wax Gourd (Benicasa hispida)", 《PLOS ONE》 * |
| BIAO JIANG等: "High-density genetic map construction and gene mapping of pericarp color in wax gourd using specific-locus amplified fragment (SLAF) sequencing", 《BMC GENOMICS》 * |
| 张玉芳等: "基因表达研究中内参基因的选择与应用", 《植物生理学报》 * |
| 牙库甫江•阿西木等: "植物基因表达转录分析中内参基因的选择与应用", 《生物技术通报》 * |
| 胡瑞波等: "植物实时荧光定量PCR内参基因的选择", 《中国农业科技导报》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110016518A (en) * | 2019-04-28 | 2019-07-16 | 福建省农业科学院作物研究所 | A kind of okra reference gene EF-1 α and its application |
| CN111575401A (en) * | 2020-07-07 | 2020-08-25 | 福建省农业科学院作物研究所 | Primer of towel gourd reference gene UBQ and application |
| CN111676230A (en) * | 2020-07-07 | 2020-09-18 | 福建省农业科学院作物研究所 | Towel gourd reference gene EF-1 alpha, and primer and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108977514B (en) | 2021-06-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108977514A (en) | For screening the primer of wax gourd real-time fluorescence quantitative PCR reference gene EF-1 α | |
| CN106636342B (en) | Acquisition method and application of EST-SSR labeled primer group developed based on ligusticum wallichii transcriptome sequence | |
| CN109022548A (en) | For screening the primer of wax gourd real-time fluorescence quantitative PCR reference gene TUA | |
| CN107190062A (en) | The screening and application of pear fruit different development stage fluorescent quantitation reference gene | |
| CN108486131A (en) | A kind of american pumpkin TUA genes and application | |
| CN113151567B (en) | SSR molecular marker and method for identifying Lepista sordida N006# strain | |
| CN113308565A (en) | Molecular marker primer combination for rapidly identifying morphological characters of waxberry leaves and application thereof | |
| CN109022547A (en) | For screening the primer of wax gourd real-time fluorescence quantitative PCR reference gene UBQ | |
| CN116179752B (en) | Primer group for identifying toxic mushroom species and PCR detection method | |
| CN104762403A (en) | GmDGK7 gene molecular marker significantly related with soybean oil content and application of GmDGK7 gene molecular marker | |
| CN108728453A (en) | A kind of giant pumpkin EF1- α genes and its application | |
| CN112239779A (en) | Primer and kit for quickly identifying sex of egg-shaped pompano and application of primer and kit | |
| CN114292901B (en) | Screening method of blueberry real-time fluorescence quantitative PCR reference genes | |
| Pérez-Palacios et al. | Deciphering the early mouse embryo transcriptome by low-input RNA-Seq | |
| CN109609512A (en) | Application of the iris PP2A gene as reference gene | |
| CN112695124B (en) | Phalaenopsis SSR molecular marker primer composition and application thereof | |
| CN103725777A (en) | Real-time fluorescence PCR (Polymerase Chain Reaction) method for rapidly detecting transgenic soybean MON89788 | |
| CN115873982B (en) | Internal reference gene for gene expression in peony stem development, primer and application thereof | |
| CN107653250B (en) | Acer palmatum reference gene and application thereof | |
| CN112725426B (en) | Specific SCAR molecular marker for sex identification of cantaloupe, primer group, kit, identification method and application of specific SCAR molecular marker | |
| CN108998566B (en) | Molecular marker primer for identifying super-red kiwi variety and application thereof | |
| KR102623818B1 (en) | Molecular marker based on chloroplast genome sequence for discriminating Prunus mume and Prunus armeniaca and uses thereof | |
| CN109456980B (en) | Bletilla striata high-temperature internal reference genes Bs18S rRNA and BUBI | |
| CN116064576B (en) | Internal reference gene of oat under ultraviolet stress, primer sequence and application of internal reference gene | |
| KR20120060790A (en) | Bio-markers for detecting tissue-specific expression in bovine's muscle regions, primer sets for detecting the bio-markers and methods of identification of bovines' muscle regions using the same |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |