CN107586860A - Three reference genes and its universal primer for being suitable to octopus category animal qPCR - Google Patents
Three reference genes and its universal primer for being suitable to octopus category animal qPCR Download PDFInfo
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- CN107586860A CN107586860A CN201711072117.0A CN201711072117A CN107586860A CN 107586860 A CN107586860 A CN 107586860A CN 201711072117 A CN201711072117 A CN 201711072117A CN 107586860 A CN107586860 A CN 107586860A
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Abstract
The invention discloses three applications suitable for octopus category animal qPCR reference gene and by the gene in real-time fluorescence quantitative PCR.Have the beneficial effect that：The gene can be stablized under varying environment and experimental condition expresses, and is not influenceed by any internal and external factor；With elongation factor 1 alpha, 18s and ubiquitin, the alpha of elongation factor 1 are reference gene, applied to real-time fluorescence quantitative PCR, the horizontal requirement of real-time fluorescence quantitative PCR detection octopus category animal gene transcriptional expression is disclosure satisfy that, improves the stability and reliability of the analysis and research of octopus category Gene Expression in Animals.
Description
Technical field
The invention belongs to field of molecular biotechnology, more particularly, to three suitable for octopus category animal qPCR reference genes and
Its universal primer.
Background technology
Suitable reference gene, it is the accurate expression pattern of identifying purpose gene in real-time fluorescence quantitative PCR (qRT-PCR)
Key factor.Preferable reference gene should possess following characteristics：Have expression in a organized way and in cell type, and not by outer
Boundary's condition influences；It is similar to destination gene expression level.Siphonopods is just to be able to widely studied marine organisms thing recent years
Kind, the research of its functional gene is also like a raging fire, but so far the reference gene of siphonopods quantitative fluorescent PCR more indiscriminately imitate shellfish etc. its
Its marine organisms, still without the reference gene suitable for siphonopods self-characteristic.
The content of the invention
Present invention aims at three reference genes for being suitable to octopus category animal qPCR are provided, the gene is in varying environment and examination
It can stablize under the conditions of testing and express, and not influenceed by any internal and external factor.
Another object of the present invention is to provide the reference gene for being suitable to octopus category animal qPCR in real-time fluorescence quantitative PCR
Application, with elongation factor-1 alpha, 18s and ubiquitin, elongation factor-1 alpha
For reference gene, the horizontal requirement of real-time fluorescence quantitative PCR detection octopus category animal gene transcriptional expression is disclosure satisfy that, improves octopus
Belong to stability, reliability and the efficiency of Gene Expression in Animals analysis and research.
The present invention is directed to the problem of being mentioned in background technology, and the technical scheme taken is：Suitable in octopus category animal qPCR
Join application of the gene in real-time fluorescence quantitative PCR, extract octopus category animal total serum IgE, reverse transcription synthesis cDNA, to the cDNA of synthesis
Carry out real-time fluorescence quantitative PCR analysis detection；Reference gene used be elongation factor-1 alpha, 18s and
Ubiquitin, elongation factor-1 alpha nucleotide sequence are as shown in SEQ ID NO.1,18s nucleotides
Sequence is as shown in SEQ ID NO.2, and ubiquitin nucleotide sequence is as shown in SEQ ID NO.3.
Preferably, in real-time fluorescence quantitative PCR analysis, the upper of elongation factor-1 alpha genes is expanded
Trip primer sequence is GTAGAGATGCACCACGAGTCACTT (as shown in SEQ ID NO.4), and downstream primer sequence is
CATACCCACGACGAAGATCCTT (as shown in SEQ ID NO.5)；Amplification 18s genes upstream primer sequence be
GCGTTCGCTTCGATGAC (as shown in SEQ ID NO.6), downstream primer sequence are GCCCTTCCGTCAATTCC (such as SEQ ID
Shown in NO.7)；The upstream primer sequence for expanding ubiquitin genes is AGAAGGTTAAGTTGGCGGTTTTG (such as SEQ ID
Shown in NO.8), downstream primer sequence is CCAGCTCCACATTCCTCGTT (as shown in SEQ ID NO.9).Upstream and downstream primer is grown
Degree is appropriate, the sequence close complementary with template, is difficult to form stable dimer and hairpin structure between primer and primer, in mistake
The efficiency of initiation of coordination site is extremely low.
Preferably, real-time fluorescence quantitative PCR amplification system is：Take 12.5 μ L SYBR green PCR master
Each 0.5 μ L mixing of mix, 1 μ L templates cDNA, upstream and downstream primer (10 μm of ol/L), adds aqua sterilisa to 25 μ L；Amplification condition is：
Mixture is followed in 95 DEG C of pre-degeneration 10min, then 95 DEG C of denaturation 15s, 60 DEG C of 1min that anneal, 72 DEG C of extension 20s, progress 40
Ring, last 72 DEG C of extensions 1min.Wherein, SYBR green PCR master mix used buy public in silent your science and technology of winged generation of match
Department.Under the conditions of being somebody's turn to do, target sequence denaturation is thorough, and it does not influence the activity of polymerase, and primer extend is complete, can reach effective expansion
Increment, and non-specific amplification is few.
Compared with prior art, the advantage of the invention is that：The invention provides three to be suitable in octopus category animal qPCR
Join gene, the gene can be stablized under varying environment and experimental condition expresses, and is not influenceed by any internal and external factor；This hair
Application of the bright reference gene additionally provided suitable for octopus category animal qPCR in real-time fluorescence quantitative PCR, with elongation
Factor-1 alpha, 18s and ubiquitin, elongation factor-1 alpha are reference gene, be disclosure satisfy that
The horizontal requirement of real-time fluorescence quantitative PCR detection octopus category animal gene transcriptional expression, improves the analysis of octopus category Gene Expression in Animals
The stability and reliability of research.
Embodiment
Below by embodiment, the present invention will be further described：
Embodiment 1：Application of the reference gene in real-time fluorescence quantitative PCR suitable for octopus category animal qPCR：
1) octopus category animal total serum IgE is extracted according to the operating instruction of Omega total RNA extraction reagent boxes, according still further to TaKaRa companies
The operating instruction synthesis cDNA of PrimeScript TM RT reagent kit with gDNA Eraser reverse transcription reagent box；
2) quantitative fluorescent PCR analysis detection is carried out to the cDNA of step 1) synthesis, reference gene used is elongation
Factor-1 alpha, 18s and ubiquitin, elongation factor-1 alpha nucleotide sequence such as SEQ ID
Shown in NO.1,18s nucleotide sequence is as shown in SEQ ID NO.2, ubiquitin nucleotide sequence such as SEQ ID NO.3
It is shown；
Amplification elongation factor-1 alpha genes upstream primer sequence be
GTAGAGATGCACCACGAGTCACTT, downstream primer sequence CATACCCACGACGAAGATCCTT；Expand the upper of 18s genes
Trip primer sequence is GCGTTCGCTTCGATGAC, downstream primer sequence GCCCTTCCGTCAATTCC；Expand ubiquitin bases
The upstream primer sequence of cause is AGAAGGTTAAGTTGGCGGTTTTG, downstream primer sequence CCAGCTCCACATTCCTCGTT；
PCR amplification system is：Take 12.5 μ L SYBR green PCR master mix, 1 μ L templates cDNA, upstream and downstream primer
(10 μm of ol/L) each 0.5 μ L mixing, adds aqua sterilisa to 25 μ L；Amplification condition is：By mixture in 95 DEG C of pre-degeneration 10min,
Then 95 DEG C of denaturation 15s, 60 DEG C of annealing 1min, 72 DEG C of extension 20s, carry out 40 circulations, last 72 DEG C of extensions 1min；
Elongation factor-1 alpha nucleotide sequence SEQ ID NO.1 are：
CCTTGGTACGAAGGCTGGGAAATTGAGAGGAAAGAGGGTAACTGCAAGGGCAATACCCTCTTCAATGCCTTGGATTC
TATTATTCCGCCGCAGAGACCAACGGAGAGACCATTGAGATTGCCCCTGCAGGATGTCTACAAGATAGGTGGTATTG
GTACCGTCCCTGTTGGCAGAGTCGAGACCGGTGTCTTGAAGCCTGGTACCGTCGTGACATTTGCACCTGCCATGGTA
TCCACTGAGGTTAGTTGATTTTTATTGTTAACTGTATTTTTTGCTTGGAGTTGTTGCTTATAGAATTTTACTTACAT
TTGATTTAATTTGTATGTTTTGCTTAGGTGAAGTCTGTAGAGATGCACCACGAGTCACTTCCAGAAGCTAACCCAGG
AGACAACGTTGGTTTTAACGTTAAGAACGTATCTGTAAAGGATCTTCGTCGTGGGTATGTGGCTGGTGACAGCAAGA
ACGACCCTCCTAAGGAAACGAAGTGCTTTGATGCCCAGGTATGTAACAAGTATTTATGAGCTCGTCTTAAGTTGAAC
GTGATTGTGTTTAATTTTTCTCTAAGTATATTCTATTCTCTCATGTAACTTATCATTCTCCTTACAGGTTATTGTCA
TTAACCACCCT；
18s nucleotide sequence SEQ ID NO.2 are：
CGTTTTCCTCGATCAAGAGCGAAAGTCGGAGGTTCGAAGACGATCAGATACCGTCGTAGTTCCGACCGTAAACGATG
CCGACTGGCGCTCCGCCGGCGTTCGCTTCGATGACCCGGCGGGGAGCCTGCGCGGGAAACCATAGTCGGTTCCGGGG
GGAGTATGGTTGCAAAACAGAAACTTAAAGGAATTGACGGAAGGGCACCACCAGGAGTGGAGCCTGCGGCTTAATTT
GACCCAACACGGGAAAACTCACCCGGTCCGGACACCGGTAGGATTGACAGATCGATAGCTCTTTCTCGATTCGGTGG
GTGGTGGTGCATGGCCGTTCTTAGTTGGTGGAGCGATCCGTCTGGTTGATTCCGATAACGAACGAGACTCCGGCCTG
CTAACTAGCGCGTCGATTAGTGCTTTTCCGGTAGCGT；
Ubiquitin nucleotide sequence SEQ ID NO.3 are：
TGTCAAGGCAAAGATTCAAGATAAGGAGGGAATTCCTCCAGATCAGCAAAGATTGATCTTTGCTGGTAAGCAGCTTG
AGGATGGCCGTACACTGTCAGACTACAATATTCAAAAGGAATCAACCCTTCATTTGGTCCTTCGTCTGCGAGGTGGC
GCCAAGAAGCGCAAGAAGAAGAATTACACCACTCCCAAAAAGAATAAGCATAAGAAGAAGAAGGTTAAGTTGGCGGT
TTTGAAATATTACAAGGTTGATGAAAATGGTAAAATCACTCGTTTGCGTCGTGAGTGTCCCAACGAGGAATGTGGAG
CTGGTGTGTTTATGGC。
Embodiment 2：Application of the reference gene in real-time fluorescence quantitative PCR suitable for octopus category animal qPCR：
1) octopus category animal total serum IgE is extracted according to the operating instruction of Omega total RNA extraction reagent boxes, according still further to TaKaRa companies
The operating instruction synthesis cDNA of PrimeScript TM RT reagent kit with gDNA Eraser reverse transcription reagent box；
2) quantitative fluorescent PCR analysis detection is carried out to the cDNA of step 1) synthesis, reference gene used is elongation
Factor-1 alpha, 18s and ubiquitin, elongation factor-1 alpha nucleotide sequence such as SEQ ID
Shown in NO.1,18s nucleotide sequence is as shown in SEQ ID NO.2, ubiquitin nucleotide sequence such as SEQ ID NO.3
It is shown；Amplification elongation factor-1 alpha genes upstream primer sequence be
GTAGAGATGCACCACGAGTCACTT, downstream primer sequence CATACCCACGACGAAGATCCTT；Expand the upper of 18s genes
Trip primer sequence is GCGTTCGCTTCGATGAC, downstream primer sequence GCCCTTCCGTCAATTCC；Expand ubiquitin bases
The upstream primer sequence of cause is AGAAGGTTAAGTTGGCGGTTTTG, downstream primer sequence CCAGCTCCACATTCCTCGTT；
PCR amplification system is：Take 12.5 μ L SYBR green PCR master mix, 1 μ L templates cDNA, upstream and downstream primer (10 μ
Mol/L) each 0.5 μ L mixing, adds the chloro mandelic acids of 1ng tetra-, 2.3ng2- chloro-5-nitrobenzoic acids, adds aqua sterilisa to 25 μ
L；Amplification condition is：By mixture in 95 DEG C of pre-degeneration 10min, then 95 DEG C of denaturation 15s, 60 DEG C of annealing 1min, 72 DEG C of extensions
10s, carry out 40 circulations, last 72 DEG C of extensions 1min.Four chloro mandelic acids, 2- chloro-5-nitrobenzoic acids and the SYBR added
Buffer solution in green PCR master mix has synergy, makes to polymerize in SYBR green PCR master mix
Enzyme is mutated in D452k sites, improves the activity of enzyme, increases the Drawing rate of genomic DNA, and it is dynamic to improve octopus category
The efficiency of thing gene expression analysis research.
Embodiment of above is merely to illustrate the present invention, and not limitation of the present invention, the ordinary skill people of this area
Member, without departing from the spirit and scope of the present invention, it can also make a variety of changes and modification.Therefore, it is all equivalent
Technical scheme fall within scope of the invention, scope of patent protection of the invention should be defined by the claims.
Sequence table
<110>Zhejiang Ocean university
<120>Three reference genes and its universal primer for being suitable to octopus category animal qPCR
<160> 9
<170> SIPOSequenceListing 1.0
<210> 1
<211> 627
<212> DNA
<213>True octopus (Octopus vulgaris)
<400> 1
ccttggtacg aaggctggga aattgagagg aaagagggta actgcaaggg caataccctc 60
ttcaatgcct tggattctat tattccgccg cagagaccaa cggagagacc attgagattg 120
cccctgcagg atgtctacaa gataggtggt attggtaccg tccctgttgg cagagtcgag 180
accggtgtct tgaagcctgg taccgtcgtg acatttgcac ctgccatggt atccactgag 240
gttagttgat ttttattgtt aactgtattt tttgcttgga gttgttgctt atagaatttt 300
acttacattt gatttaattt gtatgttttg cttaggtgaa gtctgtagag atgcaccacg 360
agtcacttcc agaagctaac ccaggagaca acgttggttt taacgttaag aacgtatctg 420
taaaggatct tcgtcgtggg tatgtggctg gtgacagcaa gaacgaccct cctaaggaaa 480
cgaagtgctt tgatgcccag gtatgtaaca agtatttatg agctcgtctt aagttgaacg 540
tgattgtgtt taatttttct ctaagtatat tctattctct catgtaactt atcattctcc 600
ttacaggtta ttgtcattaa ccaccct 627
<210> 2
<211> 422
<212> DNA
<213>True octopus (Octopus vulgaris)
<400> 2
cgttttcctc gatcaagagc gaaagtcgga ggttcgaaga cgatcagata ccgtcgtagt 60
tccgaccgta aacgatgccg actggcgctc cgccggcgtt cgcttcgatg acccggcggg 120
gagcctgcgc gggaaaccat agtcggttcc ggggggagta tggttgcaaa acagaaactt 180
aaaggaattg acggaagggc accaccagga gtggagcctg cggcttaatt tgacccaaca 240
cgggaaaact cacccggtcc ggacaccggt aggattgaca gatcgatagc tctttctcga 300
ttcggtgggt ggtggtgcat ggccgttctt agttggtgga gcgatccgtc tggttgattc 360
cgataacgaa cgagactccg gcctgctaac tagcgcgtcg attagtgctt ttccggtagc 420
gt 422
<210> 3
<211> 324
<212> DNA
<213>True octopus (Octopus vulgaris)
<400> 3
tgtcaaggca aagattcaag ataaggaggg aattcctcca gatcagcaaa gattgatctt 60
tgctggtaag cagcttgagg atggccgtac actgtcagac tacaatattc aaaaggaatc 120
aacccttcat ttggtccttc gtctgcgagg tggcgccaag aagcgcaaga agaagaatta 180
caccactccc aaaaagaata agcataagaa gaagaaggtt aagttggcgg ttttgaaata 240
ttacaaggtt gatgaaaatg gtaaaatcac tcgtttgcgt cgtgagtgtc ccaacgagga 300
atgtggagct ggtgtgttta tggc 324
<210> 4
<211> 24
<212> DNA
<213>Artificial synthesized (Saccharum)
<400> 4
gtagagatgc accacgagtc actt 24
<210> 5
<211> 22
<212> DNA
<213>Artificial synthesized (Saccharum)
<400> 5
catacccacg acgaagatcc tt 22
<210> 6
<211> 17
<212> DNA
<213>Artificial synthesized (Saccharum)
<400> 6
gcgttcgctt cgatgac 17
<210> 7
<211> 17
<212> DNA
<213>Artificial synthesized (Saccharum)
<400> 7
gcccttccgt caattcc 17
<210> 8
<211> 23
<212> DNA
<213>Artificial synthesized (Saccharum)
<400> 8
agaaggttaa gttggcggtt ttg 23
<210> 9
<211> 20
<212> DNA
<213>Artificial synthesized (Saccharum)
<400> 9
ccagctccac attcctcgtt 20
Claims (6)
1. the reference gene suitable for octopus category animal qPCR, it is characterised in that：The reference gene is elongation factor-1
Alpha, 18s and ubiquitin, elongation factor-1 alpha nucleotide sequence SEQ ID NO.1 are：
CCTTGGTACGAAGGCTGGGAAATTGAGAGGAAAGAGGGTAACTGCAAGGGCAATACCCTCTTCAATGCCTTGGATTC
TATTATTCCGCCGCAGAGACCAACGGAGAGACCATTGAGATTGCCCCTGCAGGATGTCTACAAGATAGGTGGTATTG
GTACCGTCCCTGTTGGCAGAGTCGAGACCGGTGTCTTGAAGCCTGGTACCGTCGTGACATTTGCACCTGCCATGGTA
TCCACTGAGGTTAGTTGATTTTTATTGTTAACTGTATTTTTTGCTTGGAGTTGTTGCTTATAGAATTTTACTTACAT
TTGATTTAATTTGTATGTTTTGCTTAGGTGAAGTCTGTAGAGATGCACCACGAGTCACTTCCAGAAGCTAACCCAGG
AGACAACGTTGGTTTTAACGTTAAGAACGTATCTGTAAAGGATCTTCGTCGTGGGTATGTGGCTGGTGACAGCAAGA
ACGACCCTCCTAAGGAAACGAAGTGCTTTGATGCCCAGGTATGTAACAAGTATTTATGAGCTCGTCTTAAGTTGAAC
GTGATTGTGTTTAATTTTTCTCTAAGTATATTCTATTCTCTCATGTAACTTATCATTCTCCTTACAGGTTATTGTCA
TTAACCACCCT；18s nucleotide sequence SEQ ID NO.2 are：CGTTTTCCTCGATCAAGAGCGAAAGTCGGAGGTTC
GAAGACGATCAGATACCGTCGTAGTTCCGACCGTAAACGATGCCGACTGGCGCTCCGCCGGCGTTCGCTTCGATGAC
CCGGCGGGGAGCCTGCGCGGGAAACCATAGTCGGTTCCGGGGGGAGTATGGTTGCAAAACAGAAACTTAAAGGAATT
GACGGAAGGGCACCACCAGGAGTGGAGCCTGCGGCTTAATTTGACCCAACACGGGAAAACTCACCCGGTCCGGACAC
CGGTAGGATTGACAGATCGATAGCTCTTTCTCGATTCGGTGGGTGGTGGTGCATGGCCGTTCTTAGTTGGTGGAGCG
ATCCGTCTGGTTGATTCCGATAACGAACGAGACTCCGGCCTGCTAACTAGCGCGTCGATTAGTGCTTTTCCGGTAGC
GT；Ubiquitin nucleotide sequence SEQ ID NO.3 are：TGTCAAGGCAAAGATTCAAGATAAGGAGGGAATTCCTCC
AGATCAGCAAAGATTGATCTTTGCTGGTAAGCAGCTTGAGGATGGCCGTACACTGTCAGACTACAATATTCAAAAGG
AATCAACCCTTCATTTGGTCCTTCGTCTGCGAGGTGGCGCCAAGAAGCGCAAGAAGAAGAATTACACCACTCCCAAA
AAGAATAAGCATAAGAAGAAGAAGGTTAAGTTGGCGGTTTTGAAATATTACAAGGTTGATGAAAATGGTAAAATCAC
TCGTTTGCGTCGTGAGTGTCCCAACGAGGAATGTGGAGCTGGTGTGTTTATGGC。
2. application of the reference gene suitable for octopus category animal qPCR in real-time fluorescence quantitative PCR, it is characterised in that：In described
It is elongation factor-1 alpha, 18s and ubiquitin, elongation factor-1 alpha to join gene
Nucleotide sequence as shown in SEQ ID NO.1,18s nucleotide sequence is as shown in SEQ ID NO.2, ubiquitin core
Nucleotide sequence is as shown in SEQ ID NO.3.
3. the application according to claim 2 suitable for octopus category animal qPCR reference gene in real-time fluorescence quantitative PCR,
It is characterized in that：The application process is：Octopus category animal total serum IgE is extracted, reverse transcription synthesis cDNA, the cDNA of synthesis is carried out in fact
When quantitative fluorescent PCR analyze.
4. it is suitable to application of the octopus category animal qPCR reference gene in real-time fluorescence quantitative PCR according to claim 3,
It is characterized in that：In the real-time fluorescence quantitative PCR analysis, the upstream of elongation factor-1 alpha genes is expanded
Primer sequence is GTAGAGATGCACCACGAGTCACTT, downstream primer sequence CATACCCACGACGAAGATCCTT；Amplification
The upstream primer sequence of 18s genes is GCGTTCGCTTCGATGAC, downstream primer sequence GCCCTTCCGTCAATTCC；Amplification
The upstream primer sequence of ubiquitin genes is AGAAGGTTAAGTTGGCGGTTTTG, and downstream primer sequence is
CCAGCTCCACATTCCTCGTT。
5. it is suitable to application of the octopus category animal qPCR reference gene in real-time fluorescence quantitative PCR according to claim 3,
It is characterized in that：The real-time fluorescence quantitative PCR amplification system is：Take 12.5 μ L SYBR green PCR master mix, 1
Each 0.5 μ L mixing of μ L templates cDNA, upstream and downstream primer (10 μm of ol/L), adds aqua sterilisa to 25 μ L.
6. it is suitable to application of the octopus category animal qPCR reference gene in real-time fluorescence quantitative PCR according to claim 3, its
It is characterised by：The real-time fluorescence quantitative PCR amplification condition is：By mixture in 95 DEG C of pre-degeneration 10min, then 95 DEG C of denaturation
15s, 60 DEG C of annealing 1min, 72 DEG C of extension 20s, carries out 40 circulations, last 72 DEG C of extensions 1min.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109055384A (en) * | 2018-07-19 | 2018-12-21 | 华侨大学 | A kind of Sipunculus nudus beta-actin gene and its application |
| CN109055507A (en) * | 2018-09-19 | 2018-12-21 | 福建省农业科学院植物保护研究所 | Detect the primer and method of Agasicles hygrophila EF gene expression characteristics |
| CN109112220A (en) * | 2018-10-22 | 2019-01-01 | 福建省农业科学院植物保护研究所 | Detect the primer and method of Agasicles hygrophila OBP gene transcription level |
| CN109136393A (en) * | 2018-10-22 | 2019-01-04 | 福建省农业科学院植物保护研究所 | Detect the primer and method of Agasicles hygrophila CCO gene expression characteristics |
| CN109182548A (en) * | 2018-10-22 | 2019-01-11 | 福建省农业科学院农业质量标准与检测技术研究所 | Detect the primer and method of Agasicles hygrophila GR gene expression characteristics |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102321758A (en) * | 2011-08-19 | 2012-01-18 | 中国科学院新疆生态与地理研究所 | Method for screening real-time fluorescent quantitative PCR internal reference molecules of desert plant Eremosparton songoricum (Litv.) Vass. |
-
2017
- 2017-11-03 CN CN201711072117.0A patent/CN107586860A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102321758A (en) * | 2011-08-19 | 2012-01-18 | 中国科学院新疆生态与地理研究所 | Method for screening real-time fluorescent quantitative PCR internal reference molecules of desert plant Eremosparton songoricum (Litv.) Vass. |
Non-Patent Citations (3)
| Title |
|---|
| GENBANK: "AY651883.1", 《NCBI》 * |
| GENBANK: "FJ617439.1", 《NCBI》 * |
| GENBANK: "FJ617440.1", 《NCBI》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109055384A (en) * | 2018-07-19 | 2018-12-21 | 华侨大学 | A kind of Sipunculus nudus beta-actin gene and its application |
| CN109055507A (en) * | 2018-09-19 | 2018-12-21 | 福建省农业科学院植物保护研究所 | Detect the primer and method of Agasicles hygrophila EF gene expression characteristics |
| CN109112220A (en) * | 2018-10-22 | 2019-01-01 | 福建省农业科学院植物保护研究所 | Detect the primer and method of Agasicles hygrophila OBP gene transcription level |
| CN109136393A (en) * | 2018-10-22 | 2019-01-04 | 福建省农业科学院植物保护研究所 | Detect the primer and method of Agasicles hygrophila CCO gene expression characteristics |
| CN109182548A (en) * | 2018-10-22 | 2019-01-11 | 福建省农业科学院农业质量标准与检测技术研究所 | Detect the primer and method of Agasicles hygrophila GR gene expression characteristics |
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