CN108384884A - One group of corn SSR molecular marker and its application - Google Patents
One group of corn SSR molecular marker and its application Download PDFInfo
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- CN108384884A CN108384884A CN201810488914.5A CN201810488914A CN108384884A CN 108384884 A CN108384884 A CN 108384884A CN 201810488914 A CN201810488914 A CN 201810488914A CN 108384884 A CN108384884 A CN 108384884A
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Abstract
The present invention provides one group of corn SSR molecular marker and its application, one group of corn SSR molecular marker totally 17, the present invention also provides the primer combinations of a set of detection SSR molecular marker suitable for Capillary Electrophoresis platform.It can be with using this 17 SSR molecular markers：(1) Maize SSR fingerprint database is built；(2) the maternal Source Tracing of corn sample；(3) corn variety identification and analysis of genetic diversity；(4) it distinguishes corn dolantin Asia 1 and steps on extra large 18 kinds.The application of corn SSR molecular marker through the invention can expand the range in corn serviceable indicia site in genomic level；For corn variety, germplasm identification, affiliation evaluation, the researchs such as cytoplasmic inheritance characteristic provide new tool, and application prospect is good.
Description
Technical field
The invention belongs to crops technical field of molecular biology, are examined for Capillary Electrophoresis specifically, being related to one group
The corn SSR molecular marker of survey technology and its application.
Background technology
Corn Purity and kind differentiation are the key links in corn seed quality control system.Corn purity at present
Identification and kind differentiating method have Morphological Identification, isodynamic enzyme or seed storage protein electrophoretic techniques etc..Wherein, the storage of seeds egg
White matter electrophoretic techniques have many advantages, such as it is simple, quick, accurate, at low cost, according to the technology formulate Its Relevant Technology Standards
It is used widely.However, since seed storage protein polymorphism is poor between the corn variety of part or bands of a spectrum show as incomplementarity class
The reasons such as type are difficult to find that characteristic strip using seed storage protein electrophoretic techniques so that part corn lacks quickly, reliably
Identification of means.
The appearance of DNA molecular marker technology provides new means for corn variety identification.Wherein, SSR marker is due to tool
There is quantity to enrich, polymorphism is high, be in genetically codominance, amplification is stablized, primer sequence is easy to be weighed extensively the features such as exchange
Depending on.The key of cultivar identification is carried out using simple repeated sequence (Simple Sequence Repeats, SSR) labelling technique
First, amplified fragments detection technique.Existing research is mostly examined using more complicated polyacrylamide gel electrophoresis combination silver staining
It surveys.But the technology is low due to resolution ratio, it is difficult to clearly distinguish the difference of several bases between amplified fragments, and to instrument and equipment and people
Member's skill set requirements are relatively high, are unfavorable for popularizing.And the SSR molecular marker technology based on capillary electrophoresis detection technology then can be with
Obtain quantitative DNA fragmentation analysis data.Compared with conventional polyacrylamide gel electrophoresis detection method, result is more smart
Really, sensitive, efficient, it is more suitable for the detection and analysis of high-volume kind.
Invention content
The object of the present invention is to provide one group of corn SSR molecular markers suitable for capillary electrophoresis detection technology.
In order to realize the object of the invention, the present invention by collect derive from a wealth of sources, phenotype and genotype are abundant, representative strong
Corn material is sequenced and is compared to the Maize genome of respective material.The present invention provides one group to be suitable for capillary electricity
Swim detection technique corn SSR molecular marker, the molecular labeling be following 17 SSR molecular markers one or more, 17
A SSR molecular marker is respectively PHI015, PHI299852, UMC1245, UMC1632, UMC1134, NC030, PHI109275,
PHI072, UMC2214, UMC1073, MMMC0481, UMC1066, PHI024, UMC1639, UMC1155, PHI053,
PHI227562。
17 SSR molecular markers pass sequentially through following primer amplification and obtain respectively：SEQ ID NO.1-2, SEQ ID
NO.3-4, SEQ ID NO.5-6, SEQ ID NO.7-8, SEQ ID NO.9-10, SEQ ID NO.11-12, SEQ ID
NO.13-14, SEQ ID NO.15-16, SEQ ID NO.17-18, SEQ ID NO.19-20, SEQ ID NO.21-22, SEQ
ID NO.23-24, SEQ ID NO.25-26, SEQ ID NO.27-28, SEQ ID NO.29-30, SEQ ID NO.31-32,
SEQ ID NO.33-34。
Further, the present invention provides the specific primers pair for expanding above-mentioned SSR molecular marker, are following any
It is right：SEQ ID NO.1-2, SEQ ID NO.3-4, SEQ ID NO.5-6, SEQ ID NO.7-8, SEQ ID NO.9-10, SEQ
ID NO.11-12, SEQ ID NO.13-14, SEQ ID NO.15-16, SEQ ID NO.17-18, SEQ ID NO.19-20,
SEQ ID NO.21-22, SEQ ID NO.23-24, SEQ ID NO.25-26, SEQ ID NO.27-28, SEQ ID NO.29-
30, SEQ ID NO.31-32, SEQ ID NO.33-34.
The present invention provides application of the above-mentioned corn SSR molecular marker in building Maize DNA Fingerprint Database.
The present invention provides application of the above-mentioned corn SSR molecular marker in corn germ plasm resource analysis of genetic diversity.
The present invention provides application of the above-mentioned corn SSR molecular marker in corn identification.
The present invention provides application of the above-mentioned corn SSR molecular marker in corn molecular mark.
The present invention provides application of the above-mentioned corn SSR molecular marker in preparing Maize genome chip.
The present invention provides the Maize genome chips containing above-mentioned corn SSR molecular marker.
The present invention provides above-mentioned corn SSR molecular markers to distinguish corn dolantin Asia 1 and step on answering in extra large 18 kinds
With.
Above-described application, includes the following steps：
1) DNA of corn sample to be measured is extracted；
2) using the DNA of step 1) extraction as template, using above-mentioned SSR molecular marker, PCR amplification is carried out；
3) capillary electrophoresis system is used to detect PCR product.
In the step 2) of above application, PCR amplification uses the reaction volume of 20 μ L, contains dNTP0.25mM, forward primer, anti-
Each 0.4 μM to primer, 1.0 unit of Taq archaeal dna polymerases, 1 × PCR buffer solutions (are free of Mg2+), MgCl21.5mM, sample DNA
10-40ng.Response procedures are：94 DEG C of pre-degeneration 4min；94 DEG C denaturation 45s, 60 DEG C annealing 45s, 72 DEG C extension 45s, totally 30
Cycle；72 DEG C of extension 10min, 4 DEG C of preservations.
Further, the present invention provides for distinguishing corn dolantin Asia 1 and stepping on the kit of extra large 18 kinds, contain
For the specific primer sets of 17 corn SSR molecular markers of the present invention.Preferably, the core of the specific primer sets
Nucleotide sequence is respectively as shown in SEQ ID NO.1-34.
Above-mentioned 17 pairs of SSR primers provided by the invention can realize genotype data on fluorescent capillary electrophoresis tube platform
Acquisition.Concrete scheme is wherein one ends 5' mark fluorescent group of each pair of primer；PCR reaction systems are prepared to be added
DNA, primer, dNTP, MgCl2, Taq enzyme, Buffer；Run response procedures；Amplified production is in fluorescent capillary electrophoresis tube system
Detection；Initial data is collected using capillary electrophoresis system software kit, quiding gene type software analyzes initial data and obtains piece
The genotype data of segment length format.
Preferably, by the present invention 17 couple specificity SSR primers progress fluorochrome label, selected altogether PET, NED,
Tetra- kinds of fluorescent dyes of VIC, FAM.The PCR product of fluorescent marker ultra-pure water is diluted 30 times；Isometric above-mentioned 4 kinds are taken respectively
Solution is mixed to form mixed liquor after dilution, and 1 μ L 0.5 μ L LZ500 molecular weight internal standards of addition are drawn from mixed liquor and 8.5 μ L are gone
Formamide is added in the special deep-well plates of DNA analysis instrument；Then by it in PCR instrument 95 DEG C denaturation 5min, take out, immediately
It is placed on ice, cooling 10min or more；It is placed to after brief centrifugation 10s on DNA analysis instrument and carries out capillary electrophoresis detection.With
GeneMapper softwares analyze the initial data of collection.Software systems will be according in the position of target peak and same swimming lane
Internal standard LZ500 be compared, directly give the accurate size of target DNA fragments.
In a preferred embodiment of the embodiment of the present invention, the SSR molecular marker of FAM fluorescent marker groups is
PHI015, PHI299852, UMC1245, UMC1632；The SSR molecular marker of VIC fluorescent marker groups be UMC1134, NC030,
PHI109275, PHI072, UMC2214；The SSR molecular marker of NED fluorescent marker groups be UMC1073, MMMC0481,
UMC1066, PHI024；The SSR molecular marker of PET fluorescent marker groups is UMC1639, UMC1155, PHI053, PHI227562.
Using corn dolantin No. 1 DNA in Asia as template, four capillarys are carried out with four groups of primers of FAM, VIC, NED, PET fluorescent marker respectively
Electrophoresis tube obtains four electrophoretogram results；Again using sub- No. 1 DNA of dolantin as template, with FAM, VIC, NED, PET fluorescent marker
Whole primer mixtures carry out Capillary Electrophoresis obtain total electrophoretogram result.By total electrophoretogram result respectively with FAM, VIC,
The result of tetra- groups of fluorescent dye primer electrophoresis of NED, PET is compared (Fig. 1), it can be seen that independent in FAM, VIC, NED, PET
The purpose peak occurred on electrophoretogram can distinguish on total electrophoretogram, and the peak of each color is not interfere with each other, that is, illustrates this
The primer combination (totally 17 pairs of primers) that invention provides can be used for a Capillary Electrophoresis, and purpose band is not interfere with each other, and is easy to judge
As a result.Extra large 18DNA is stepped on using corn and carries out identical experiment as template, and identical conclusion (Fig. 2) can be obtained.
No. 1 and extra large 18DNA is stepped on as template, drawn with FAM, VIC, NED, PET fluorescent marker using dolantin is sub- respectively in embodiment
Object combination carries out disposable Capillary Electrophoresis, respectively obtains dolantin Asia 1 and steps on the Capillary Electrophoresis figure in sea 18 and be compared.
From figure 3, it can be seen that sub- No. 1 of dolantin FAM blues peak occurs in 134bp and 141bp, steps on sea 18 and go out at 131bp and 145bp
Existing FAM blues peak；There is VIC greens peak in 83bp, 99bp and 126bp in sub- No. 1 of dolantin, step on sea 18 in 86bp, 104bp and
There is VIC greens peak in 120bp；There is NED yellow peak in 136bp and 166bp in sub- No. 1 of dolantin, steps on sea 18 in 142bp and 161bp
There is NED yellow peak；There is PET red peak in 107bp and 170bp in sub- No. 1 of dolantin, steps on sea 18 and occurs in 101bp and 175bp
PET red peak.The peak that dolantin Asia 1 and each fluorescent marker for stepping on sea 18 occur is not overlapped, clear and legible.Therefore, with this
The fluorescent dye primer combination that patent of invention provides can distinguish corn dolantin Asia 1 in a Capillary Electrophoresis and step on sea 18.
The amplified production of the different fluorescent markers of the present invention can carry out electrophoresis in same swimming lane, and signal is clear, amplification
Clip size difference is apparent, can accurately calculate clip size, each DNA sample electrophoresis peak type is different, is easy to judge.With sensitive
Degree is high, resolving power is good, as a result accurately and reliably, it is efficiently quick the advantages that.It is combined with primer provided by the invention, it can be quickly and easily
No. 1 and step on corn dolantin is sub- extra large 18 kinds and distinguish, realize it is cost-effective, improve efficiency, easy to operate, result is accurate
Advantage.This group of primer provided by the invention can be used for corn fingerprint map construction, cultivar identification and analysis of genetic diversity etc., answer
It is very wide with foreground.
Description of the drawings
Figure 1A-Fig. 1 D are respectively the sub- No. 1 SSR fluorescent marker Capillary Electrophoresis figure of corn dolantin, wherein Figure 1A is that dolantin is sub-
No. 1 through FAM, VIC, NED and PET fluorescent dye primer combination electrophoresis result and the only comparison through FAM labeled primer electrophoresis results.
Figure 1B is sub- No. 1 of dolantin through FAM, VIC, NED and PET fluorescent dye primer combination electrophoresis result and only through VIC labeled primer electricity
The comparison for result of swimming.Fig. 1 C are that sub- No. 1 of dolantin combines electrophoresis result and only pass through through FAM, VIC, NED and PET fluorescent dye primer
The comparison of NED labeled primer electrophoresis results.Fig. 1 D are that sub- No. 1 of dolantin combines electricity through FAM, VIC, NED and PET fluorescent dye primer
Result of swimming and the only comparison through PET labeled primer electrophoresis results.Each figure comparison result illustrates respectively, one-color fluorescence labeled primer
The purpose peak occurred on electrophoretogram can combine in four color fluorescent dye primers and distinguish that (arrow meaning is shown as out on electrophoretogram
Existing purpose peak), and each purpose peak is not interfere with each other, and can clearly read the DNA fingerprint information of kind.Offer of the present invention is provided
Primer combination can be used for a Capillary Electrophoresis, purpose band is not interfere with each other, when being easy to save while judging result again
Between, experiment reagent and consumptive material.
Fig. 2A-Fig. 2 D are the SSR fluorescent marker capillary electrophoresis detection results that corn steps on sea 18.Wherein, Fig. 2A is to step on sea
18 through FAM, VIC, NED and PET fluorescent dye primer combination electrophoresis result and the only comparison through FAM labeled primer electrophoresis results.
Fig. 2 B are to step on sea 18 through FAM, VIC, NED and PET fluorescent dye primer combination electrophoresis result and only through VIC labeled primer electrophoresis knots
The comparison of fruit.Fig. 2 C are to step on sea 18 to combine electrophoresis result through FAM, VIC, NED and PET fluorescent dye primer and only mark through NED
The comparison of primer electrophoresis result.Fig. 2 D be step on sea 18 through FAM, VIC, NED and PET fluorescent dye primer combine electrophoresis result with only
Comparison through PET labeled primer electrophoresis results.The comparison result of each figure illustrates respectively, on the electrophoretogram of one-color fluorescence labeled primer
The purpose peak of appearance can four color fluorescent dye primers combine electrophoretogram on distinguish (arrow meaning be shown as occur purpose
Peak), and each purpose peak is not interfere with each other, and can clearly read the DNA fingerprint information of kind.Illustrate primer sets provided by the invention
Conjunction can be used for a Capillary Electrophoresis, and purpose band is not interfere with each other, and save time, experiment examination while being easy to judging result again
Agent and consumptive material.Figure 1A-Fig. 1 D and Fig. 2A-Fig. 2 D are in order to illustrate the same purpose, two kinds (dolantin sub- No. 1 and step on sea 18)
Experimental result can more illustrate this patent provide primer combination practicability and reliability.
Fig. 3 is to carry out capillary with sea 18 is stepped on for No. 1 to corn dolantin Asia with 17 corn SSR molecular markers provided by the present application
Electrophoresis tube figure, and to result that the two is compared.As a result illustrate, the peak (blue) for the FAM mesh that No. 1 (above) in dolantin Asia is shown
It is clear and legible with the peak (being marked with arrow) for the FAM mesh for stepping on extra large 18 (figure below) display, and purpose peak molecular weight is different.Similarly,
The peak molecular weight for VIC, NED, PET mesh that two kinds are shown is different and clear and legible.The primer sets that explanation is provided with this patent
Closing No. 1 sub- to corn dolantin and can step on sea 18 and carry out kind differentiation.
Specific implementation mode
The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..Unless otherwise specified, embodiment
According to conventional laboratory conditions.The content not being described in detail in present specification belongs to existing well known to professional and technical personnel in the field
There is technology.
If not specified, biochemical reagents used in the embodiment of the present invention are commercially available, and corn material used is this field
Public corn.
The determination of 1 corn SSR molecular marker of embodiment and primer
36 corn varieties are taken to carry out SSR primer screenings.With denaturing polyacrylamide gel electrophoresis, from 987 pairs of SSR primers
(https://ftp.maizegdb.org/MaizeGDB/FTP/SSRs/) select that site is amplifiable, and banding pattern is clear, polymorphism compared with
High primer is as optional primer.According to optional primer 36 kind allelics molecular weight ranges, PIC values etc., i.e.,
Each combination is made of primer as much as possible, organizes the condition that interior all primer allele ranges are not superimposed, and composition four draws
The merging of object group is respectively synthesized the primer with FAM, VIC, NED, PET fluorophor.It is shown in Table 1.It will be appreciated by those skilled in the art that
Other fluorophors including FAM, VIC, NED, PET also may be selected in the art to modify four groups of primers in table 1, only
It wants every group of inner primer to mark identical fluorophor, is not limited to select any fluorophor.
1 corn SSR Capillary Electrophoresis primer of table and primer combination
Embodiment 2 distinguishes corn variety using SSR molecular marker provided by the invention
(1) DNA rapid extractions
DNA extractions are carried out to corn seed using alkaline-heating method.The method operation is fast and convenient, without poisonous and harmful reagent, fits
Prepared by the DNA for field of plant molecular biology, and to Seed purity assessment, detection GMOs time is greatly shortened,
Detection efficiency is improved, testing cost is reduced and is of great significance.Concrete operation step is：If taking corn seed dry granular, it is placed in
In 1.5mL centrifuge tubes；400 μ L NaOH (1M) are added in centrifuge tube, it is ensured that be completely soaked seed, boiling water bath 5min；From
200 μ L Tris-HCl (1M, pH 8.0), boiling water bath 1min are added in heart pipe；200 μ L TE buffer solutions (pH 8.0) are added, fill
Divide spare after dissolving.
(2) quality and quantity of DNA sample
The OD values that DNA sample 260nm and 280nm are detected on UV detector, select OD260/280Value is 1.8-
1.9 sample is for detecting.
(3) core primers select
By analyzing distribution, polymorphism level, PCR amplification stability and the amplified production banding pattern of primer on chromosome,
The primer that disclosure satisfy that capillary detection technique of fluorescence that selection example 1 determines, is specifically shown in Table 1.
(4) capillary electrophoresis detection
The specific SSR primers filtered out are subjected to fluorochrome label, select tetra- kinds of fluorescence of PET, NED, VIC, FAM altogether
Dyestuff.The PCR product of fluorescent marker ultra-pure water is diluted 30 times；Solution mixes shape after taking isometric above-mentioned 4 kinds of dilutions respectively
At mixed liquor, 1 μ L 0.5 μ L LZ500 molecular weight internal standards of addition are drawn from mixed liquor and 8.5 μ L deionized formamides are added to
In the special deep-well plates of DNA analysis instrument；Then by it, 95 DEG C of denaturation 5min, taking-up are immediately placed on ice in PCR instrument, cooling
10min or more；It is placed to after brief centrifugation 10s on DNA analysis instrument and carries out capillary electrophoresis detection.With GeneMapper softwares pair
The initial data of collection is analyzed.Software systems are compared according to the position of target peak and the internal standard LZ500 in same swimming lane
Compared with directly giving the accurate size of target DNA fragments.Each group primer amplification corn dolantin is sub- in table 1 and steps on 18 gained of sea by No. 1
The purpose band size of amplified production is shown in Table 2.
2 corn dolantin Asia 1 of table and the difference peak band and size for stepping on sea 18
(5) judgement of primer combination
Using corn dolantin No. 1 DNA in Asia as template, carried out respectively with four groups of primers of FAM, VIC, NED, PET fluorescent marker
Four Capillary Electrophoresis obtain four electrophoretogram results；It is glimmering with FAM, VIC, NED, PET again using No. 1 DNA in dolantin Asia as template
Whole primer mixtures of signal carry out Capillary Electrophoresis and obtain total electrophoretogram result.By total electrophoretogram result respectively with FAM,
The result of tetra- groups of fluorescent dye primer electrophoresis of VIC, NED, PET is compared (Figure 1A-Fig. 1 D), it can be seen that FAM, VIC,
The purpose peak occurred on the independent electrophoretogram of NED, PET can distinguish on total electrophoretogram, and the peak of each color is not done mutually
It disturbs, that is, illustrates that primer mixture provided by the invention can be used for a Capillary Electrophoresis, purpose band is not interfere with each other, and is easy to judge
As a result.Extra large 18DNA is stepped on using corn and carries out identical experiment as template, and identical conclusion (Fig. 2A-Fig. 2 D) can be obtained.
(6) dolantin is sub- and steps on the differentiations of extra large 18 kinds by No. 1
No. 1 and extra large 18DNA is stepped on as template, drawn with FAM, VIC, NED, PET fluorescent marker using dolantin is sub- respectively in embodiment
Object combines (being shown in Table 1) and carries out disposable Capillary Electrophoresis, respectively obtains sub- No. 1 of dolantin and goes forward side by side with the Capillary Electrophoresis figure for stepping on sea 18
Row compares.From figure 3, it can be seen that there is FAM blues peak in 134bp and 141bp in sub- No. 1 of dolantin, step on sea 18 in 131bp and
Occurs FAM blues peak at 145bp；There is VIC greens peak in 83bp, 99bp and 126bp in sub- No. 1 of dolantin, step on sea 18 86bp,
There is VIC greens peak in 104bp and 120bp；There is NED yellow peak in 136bp and 166bp in sub- No. 1 of dolantin, steps on sea 18 in 142bp
There is NED yellow peak with 161bp；There is PET red peak in 107bp and 170bp in sub- No. 1 of dolantin, step on sea 18 in 101bp and
There is PET red peak in 175bp.The peak that dolantin Asia 1 and each fluorescent marker for stepping on sea 18 occur is not overlapped, clear and legible.
Therefore, the fluorescent dye primer combination (table 1) provided with patent of the present invention can distinguish corn dolantin in a Capillary Electrophoresis
Sub- No. 1 and step on sea 18.
Although above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements, belong to the scope of protection of present invention without departing from theon the basis of the spirit of the present invention.
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<110>Agricultural product quality and safety research institute of Heilongjiang Institute of Agricultural Sciences
<120>One group of corn SSR molecular marker and its application
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gcaaggagta gggaggacgt g 21
<210> 29
<211> 24
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 29
tcttttattg tgcccgttga gatt 24
<210> 30
<211> 24
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 30
cctgagggtg atttgtctgt ctct 24
<210> 31
<211> 21
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 31
tgataaagct cagccacaag g 21
<210> 32
<211> 18
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 32
atctcggcta cggccaga 18
<210> 33
<211> 25
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 33
ctgcctctca gattcagaga ttgac 25
<210> 34
<211> 20
<212> DNA
<213>Artificial sequence (Artificial Sequence)
<400> 34
aacccaacgt actccggcag 20
Claims (10)
1. the corn SSR molecular marker suitable for capillary electrophoresis detection technology, which is characterized in that the molecular labeling is following
The one or more of 17 SSR molecular markers, respectively PHI015, PHI299852, UMC1245, UMC1632, UMC1134,
NC030, PHI109275, PHI072, UMC2214, UMC1073, MMMC0481, UMC1066, PHI024, UMC1639,
UMC1155, PHI053, PHI227562.
2. corn SSR molecular marker according to claim 1, which is characterized in that 17 SSR molecular markers respectively according to
It is secondary to be obtained by following primer amplification：SEQ ID NO.1-2, SEQ ID NO.3-4, SEQ ID NO.5-6, SEQ ID NO.7-
8, SEQ ID NO.9-10, SEQ ID NO.11-12, SEQ ID NO.13-14, SEQ ID NO.15-16, SEQ ID
NO.17-18, SEQ ID NO.19-20, SEQ ID NO.21-22, SEQ ID NO.23-24, SEQ ID NO.25-26, SEQ
ID NO.27-28, SEQ ID NO.29-30, SEQ ID NO.31-32, SEQ ID NO.33-34.
3. application of the corn SSR molecular marker as claimed in claim 1 or 2 in building Maize DNA Fingerprint Database.
4. application of the corn SSR molecular marker as claimed in claim 1 or 2 in corn germ plasm resource analysis of genetic diversity.
5. application of the corn SSR molecular marker described in claims 1 or 2 in corn identification.
6. application of the corn SSR molecular marker in corn molecular mark described in claims 1 or 2.
7. the Maize genome chip containing corn SSR molecular marker described in claims 1 or 2.
8. corn SSR molecular marker described in claims 1 or 2 is distinguishing corn dolantin Asia 1 and is stepping on the application in extra large 18 kinds.
9. according to any applications of claim 3-8, which is characterized in that include the following steps：
1) DNA of corn sample to be measured is extracted；
2) using the DNA of step 1) extraction as template, according to the corn SSR molecular marker, PCR amplification is carried out；
3) capillary electrophoresis system is used to detect PCR product.
10. for differentiating corn dolantin Asia 1 and stepping on the kit of extra large 18 kinds, which is characterized in that contain and be directed to claim 1
The specific primer sets of the corn SSR molecular marker.
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| CN112167051A (en) * | 2020-09-24 | 2021-01-05 | 河南省农业科学院 | Method for creating high-lysine corn breeding material |
| CN112646921A (en) * | 2020-12-31 | 2021-04-13 | 中国农业大学 | SSR (simple sequence repeat) marker for southern rust of corn and application of SSR marker |
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| CN112646921A (en) * | 2020-12-31 | 2021-04-13 | 中国农业大学 | SSR (simple sequence repeat) marker for southern rust of corn and application of SSR marker |
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| Publication number | Publication date |
|---|---|
| CN108384884B (en) | 2021-02-09 |
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