CN1570623A - Acidic capillary electrophoresis identification method for high molecular weight glutelin subunit of wheat - Google Patents
Acidic capillary electrophoresis identification method for high molecular weight glutelin subunit of wheat Download PDFInfo
- Publication number
- CN1570623A CN1570623A CN 200410037257 CN200410037257A CN1570623A CN 1570623 A CN1570623 A CN 1570623A CN 200410037257 CN200410037257 CN 200410037257 CN 200410037257 A CN200410037257 A CN 200410037257A CN 1570623 A CN1570623 A CN 1570623A
- Authority
- CN
- China
- Prior art keywords
- capillary electrophoresis
- wheat
- 6min
- buffer
- subunit
- 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
Images
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
This invention relates to a determination method for wheat high molecular weight glutelin subunit, especially a method through technique of acid capillary electrophoresis. The key emphasis in current wheat breeding work is to improve the HMW-GS component, especially to improve the frequency of excellent high molecular weight glutelin subunit or subunit pairs The combination of current usual breeding method with label assistance selection techniques is the main means for wheat breeding improvement, wherein the rapid, microscale and accurate determination of excellent protein subunit for generation in the early hybrid time is the key to improve the breeding efficiency. The main task of this invention is to provide the buffer type and additives and condition parameter of acid capillary electrophoresis and clean-up procedures of capillary.
Description
Technical field
The present invention relates to the authentication method of wheat high-molecular-weight glutelin subunit, particularly relate to the method for wheat high-molecular-weight glutelin subunit being identified by acid capillary electrophoresis (A-CE) technology.
Background technology
The wheat seed storage protein mainly is made up of alcohol soluble protein and glutelin, and they have the heterogeneity and the complicacy of height on forming.The alcohol soluble protein molecular weight is between 30000-80000 dalton, under the acid gel deposition condition, kind is separable to go out 15-30 component, can be divided into α, β, γ and four kinds of alcohol soluble proteins of ω according to its relative mobility, they account for 25%, 30%, 30% and 15% of its total amount respectively.Glutenin comprises two kinds of high-molecular-weight glutelin subunit (HMW-GS) and low-molecular-weight glutenin subunits (LMW-GS), and they account for about 10% and 30% of endosperm total protein content respectively.
Studies have shown that in a large number the composition and the content of wheat storage protein, particularly high-molecular-weight glutelin subunit have a very important role to quality.Flour can be processed into multiple special foods such as various bread, noodles, biscuit, cake, steamed bun, its special system face characteristic mainly is by the disulfide bond polymer structures shape of storage protein, the elasticity of glutelin major decision dough wherein, alcohol soluble protein then determines the ductility of dough.Known at present that HMW-GS is by Glu-A1, Glu-B1 on the part 1 homology group chromosome, Glu-D1 site coding.All there are two close linkage genes in each site, controls the less y type subunit of bigger x type subunit of molecular weight and molecular weight (molecular weight that records with the SDS-PAGE method is 90-150KD) respectively.Because portion gene is in reticent and expression status not, most common wheat kinds only can detect 3-5 high-molecular-weight glutelin subunit on the SDS-PAGE collection of illustrative plates, wherein 2 by Glu-D1 control, and 1 or 2 are by Glu-B1 control, and 1 or 0 are controlled by Glu-A1.On each site, all there is allelic variation widely, in bread wheat, identified and named more than 20 HMW-GS allele at present.Different subunits and allele thereof are different to the effect of wheat quality, in detected high-molecular-weight glutelin subunit, and 1Dx5+1Dy10 (4), 1Bx17+1By18 (3), 1Bx7+1By8 (3) and 1Bx13+1By16 (3), 1Ax1 (3), 1Ax2
*(3) etc. subunit to or subunit relevant with wheat high-quality quality (being the quality scoring in the bracket), and 1Dx2+1Dy12 is that subunit inferior is right, subunit plays an important role to quality to 1Dx4+1Dy12, its quality is marked and is only second to 1Dx5+1Dy10.The right frequency of high quality subunits such as 1Dx5+1Dy10,1Bx17+1By18 is very low in China's commercial variety, and this may be to cause one of inferior major reason.Current emphasis in Wheat Breeding for Quality work is the improvement that HMW-GS is formed, and particularly will improve the right frequency of high-quality high-molecular weight gluten subunit or subunit.
In recent years, although demonstrated very big application prospect by genetic engineering and molecular marker assisted selection means improvement wheat quality, but these methods are expense costliness, analysis cost height, time-consuming often, and also have with a certain distance from practical application at the genetic engineering and the molecular marker-assisted selection method of high-quality storage protein gene, also be difficult to satisfy at present the requirement of quality breeding work.Therefore be badly in need of the new more reliable high-quality genetic marker assisted Selection technology of research and development.At present to combine with the marker assisted selection technology be the main means of Wheat Quality Improvement to the conventional breeding method, wherein hybridize early generation high-quality storage protein subunit fast, trace, identify it is the key of raising breeding efficiency accurately.
Summary of the invention
The purpose of this invention is to provide a kind of method of utilizing acid capillary electrophoresis technology Rapid identification wheat high-molecular-weight glutelin subunit, main task is to solve type and adding ingredient, deposition condition parameter and the capillary douche program that provides the damping fluid of acid capillary electrophoresis at the evaluation of the high-molecular-weight glutelin subunit of preserving in the wheat seed, and this method can be made wheat seed high-quality storage protein subunit at the wheat hybridizing early generation and being identified fast and accurately and screening.
Technical scheme of the present invention comprises material sampling, specimen preparation, high performance capillary electrophoresis and Data Management Analysis, what wherein material sampling, specimen preparation and data processing adopted is conventional method, electrophoretic buffer when it is characterized in that high performance capillary electrophoresis (buffer) is 100-120mm phosphoric acid-glycine buffer, this pH of buffer value is 2.5-2.8, be added with 18-22% acetonitrile and 0.04-0.06% hydroxyprolyl-methylcellulose (HPMC), concrete deposition condition parameter is:
Capillary inner diameter: 25 or 50 μ m
Capillary pipe length: 24-28cm
Voltage: 10.0-13.5KV
Temperature: 37-40 ℃
Sample introduction: 8-12KV, 6-10sec
In above-mentioned technical scheme, for can the continuous detecting sample and guarantee the accuracy of testing result, when carrying out high performance capillary electrophoresis, kapillary be washed and must operate by cleaning procedure, according to the difference of flushing purpose, cleaning procedure specifically is divided into following four kinds of situations:
1, will carry out the pre-service flushing before use for new kapillary, cleaning procedure is as follows:
H
2O 4-6min
0.1MNaOH 9-12min
H
2O 4-6min
1MH
3PO
4 8-12min
Buffer 50-75min
2, before the sample introduction kapillary is washed, cleaning procedure is as follows:
H
2O 1.5-2.5min
0.1MNaOH 4-6min
H
2O 4-6min
1MH
3PO
4 4-6min
Buffer 16-24min
3, when once identifying a plurality of sample of mensuration, will wash between twice sample detection, cleaning procedure is as follows:
1MH
3PO
4 1.5-3min
Buffer 1.5-2.5min
4, detect the end back kapillary is washed, cleaning procedure is as follows:
1MH
3PO
4 4-6min
H
2O 4-6min
0.1MNaOH 1.5-2.5min
H
2O 13-17min
N
2 9-11min
Can set up important HMW-GS standard capillary electrophoresis pattern by the present invention, and then form the complete technical system of a cover that high quality subunit is identified.
The invention has the advantages that:
(1) amount of samples is few: trace detection is the key of carrying out hybridization early generation high-quality gene Selection.Present technique only needs single seed not have embryo end 10-15mg endosperm flour, and can carry out replicate analysis more than 5 times.In addition, the protein dry sample that is extracted can keep for a long time and not influence separating effect.
(2) disengaging time is short: utilize above-mentioned electrophoresis parameter, single sample generally separates at 12min to be finished, and conventional SDS-PAGE method generally needs 1-2 days time.
(3) subunit electrophoretic resolution height, be easy to use: under the acidic buffer condition, most HMW-GS show as a main peak and 1-3 submaximum (as shown in drawings).Every meter theoretical cam curve of A-CE technology is higher than high performance liquid chromatography and SDS-PAGE method far away in theory.This technology can be separated fast and identifies the present high-quality HMW gluten subunit of identifying, comprises the subunit that some SDS-PAGE and HPLC method are difficult to distinguish, as 17+18, and 2 and 2
*The new subunit very similar with some mobilities of discovered in recent years, and 10 and 12,2 with 5 grades (RP-PLC can not effectively separate).By with the comparative analysis of high quality subunit standard diagram and single sample and compound sample, can accurately identify 5+10 and 2+12 and 14+15,13+16,17+18,7+8,1,2
*Deng subunit.
(4) good reproducibility: utilize above-mentioned purging method can obtain better repeatability and separate.The relative standard deviation of peak transit time and peak height (RSD) is respectively less than 1% and 3% between repetition, no peak conditions of streaking.
(5) separation automation degree height: Capillary Electrophoresis from sample introduction to interpretation of result automaticity height, easy easy operating, this is that traditional SDS-PAGE method is incomparable.
(6) analysis cost is low: except the Capillary Electrophoresis instrument was more expensive, analysis cost was low, generally only needed some common agents, and the equal consumption of all reagent seldom and need not use poisonous medicines such as third rare acid amides.Therefore, compare with classic method, A-CE analysis of technology cost is lower.
In addition, this authentication method also has important use value to fields such as wheat circulation and processing and the protections of kind rights and interests.
Description of drawings
Fig. 1: wheat breed Hope (1,6+8,5+10) acid capillary electrophoresis (A-CE) of high-molecular-weight glutelin subunit (HMW-GS) is separated relatively collection of illustrative plates of repeatability continuous 20 times.Show the 1st, 5,10,15 electrophoresis patterns that separate with 20 times among the figure.
Fig. 2: the acid capillary electrophoresis separating spectrum of wheat breed China spring, Hope and durum wheat kind Simeto high-molecular-weight glutelin subunit.
The A China spring (N, 7+8,2+12)
B?Hope(1，6+8，5+10)
C?Simeto(N，7+8)
Fig. 3: club wheat (Triticum compactum L., 2n=6x=42, AABBDD) Club20 (N, 17+18,2+12) (5+10) single sample and compound sample (1: 1) HMW-GS acid capillary electrophoresis is separated and is identified collection of illustrative plates for N, 17+18 with bread wheat kind Kontrast.
Embodiment
1. material sampling
As test material, the picked at random seed does not have endosperm end 10-15mg and is used for glutelin and extracts and Capillary Electrophoresis with the seed of wheat breed Hope.
2. specimen preparation
High-molecular-weight glutelin subunit (HMW-GS) extracting method:
Standby reagent:
A.50% n-propanol
B.50% n-propanol+1%DTT
C. acetone (analyzing pure)
D.20% acetonitrile+0.1% trifluoroacetic acid (TFA)
Method of operating:
(1) gets single seed, cut no endosperm end (about 10-15mg), accurately use sulphur after the weighing with blade
Acid paper and adz-eye hammer are pricked and are broken into fine powder, put into the 1.5ml centrifuge tube;
(2) add 50% n-propanol 1ml, 45 ℃ of water-baths were extracted 10 minutes, centrifugal 5 minutes of 10000rpm,
Remove supernatant;
(3) repeat above-mentioned steps three times, support by the arm the supernatant that stays and blot with filter paper;
(4) add 50% n-propanol+1%DTT 800 μ l, 1200rpm was extracted in 45 ℃ of concussions 30 minutes
Centrifugal 10 minutes, get supernatant and change in the new centrifuge tube of clean 1.5ml;
(5) add precooling and analyze pure acetone to final concentration 40%, room temperature staticly settled 30 minutes, 12000rpm
Centrifugal 15 minutes, the gained precipitation was HMW-GS;
(6) in proportion (w/v:1mg+10 μ l) adds 20% acetonitrile+0.1%w/v trifluoroacetic acid (TFA),
Fully dissolving, can carry out capillary electrophoresis analysis after centrifugal; Simultaneously also precipitation can be put into-20
Preserve in ℃, capillary electrophoresis analysis is carried out in dissolving when need waiting.
3. high performance capillary electrophoresis (HPCE)
Instrument and equipment
The high performance capillary electrophoresis system (be furnished with software and be used for system operation and data processing) that Bio-Rad (Biofocus 3000) company produces.The fused-silica capillary column internal diameter is 50 μ m, and length is 25.5cm, and skin scribbles the polyimide protective seam, and inwall is coating not all.
Standby reagent
The Capillary Electrophoresis damping fluid
(this pH of buffer value is 2.50 to 100mm phosphoric acid-glycine buffer, is added with 20% acetonitrile
With 0.05% hydroxyprolyl-methylcellulose (HPMC)); Distilled water.
The capillary column washing fluid
A.0.1MNaOH
B.1MH
3PO
4
C. distilled water
D. electrophoretic buffer
The capillary douche program
A. new kapillary preprocessor
H
2O 5min
0.1MNaOH 10min
H
2O 5min
1MH
3PO
4 10min
Buffer 60min
B. sample room cleaning procedure
1MH
3PO
4 2min
Buffer 2min
C. test cleaning procedure every day
Flushing before the sample introduction:
H
2O 2min
0.1MNaOH 5min
H
2O 5min
1MH
3PO
4 5min
Buffer 20min
D. finish the experiment flushing:
1MH
3PO
4 5min
H
2O 5min
0.1MNaOH 2min
H
2O 15min
N
2 10min
The Capillary Electrophoresis condition
Voltage: 12.5KV
Temperature: 40 ℃
Sample introduction: 10KV, 8sec
Electrode: by+to-.
Data processing
Utilize Biofocus Integrator software to carry out data processing, simultaneously the electrophoresis result data-switching is become the ASC sign indicating number, carry out the Excel Treatment Analysis.
To wheat breed Hope (1,6+8,5+10) electrophoresis pattern that obtains of continuous 20 repeated isolation of high-molecular-weight glutelin subunit is as shown in Figure 1.
1. material sampling
Use wheat breed China spring, Hope and durum wheat seed as test material respectively, the picked at random seed does not have endosperm end 10-15mg and is used for glutelin extraction and Capillary Electrophoresis.
2. specimen preparation
High-molecular-weight glutelin subunit (HMW-GS) extracting method:
The standby reagent of selecting for use is identical with embodiment 1 with method of operating, and the HMW-GS that obtains precipitation (w/v:1mg+10 μ l) in proportion adds 20% acetonitrile+0.1%w/v trifluoroacetic acid (TFA), fully dissolving, can carry out capillary electrophoresis analysis after centrifugal; Also precipitation can be put into-20 ℃ of preservations, capillary electrophoresis analysis is carried out in dissolving when need waiting.
3. high performance capillary electrophoresis (HPCE)
Instrument and equipment
The high performance capillary electrophoresis system (be furnished with software and be used for system operation and data processing) that Bio-Rad (Biofocus 3000) company produces.The fused-silica capillary column internal diameter is 50 μ m, and length is respectively 28cm, and skin scribbles the polyimide protective seam, and inwall is coating not all.
Standby reagent
The Capillary Electrophoresis damping fluid
(this pH of buffer value is 2.60 to 110mm phosphoric acid-glycine buffer, is added with 18% acetonitrile
With 0.04% hydroxyprolyl-methylcellulose (HPMC)); Distilled water.
The capillary column washing fluid
A.0.1MNaOH
B.1MH
3PO
4
C. distilled water
D. electrophoretic buffer
The capillary douche program
A. new kapillary preprocessor
H
2O 4min
0.1MNaOH 9min
H
2O 4min
1MH
3PO
4 8min
Buffer 50min
B. sample room cleaning procedure
1MH
3PO
4 1.5min
Buffer 1.5min
C. test cleaning procedure every day
Flushing before the sample introduction:
H
2O 1.5min
0.1MNaOH 4min
H
2O 4min
1MH
3PO
4 4min
Buffer 16min
D. finish the experiment flushing:
1MH
3PO
4 4min
H
2O 4min
0.1MNaOH 1.5min
H
2O 13min
N
2 9min
The Capillary Electrophoresis condition
Voltage: 10KV
Temperature: 37 ℃
Sample introduction: 8KV, 10sec
Electrode: by+to-.
Data processing
Utilize Biofocus Integrator software to carry out data processing, simultaneously the electrophoresis result data-switching is become the ASC sign indicating number, carry out the Excel Treatment Analysis.
The acid capillary electrophoresis separating spectrum of China spring, Hope and durum wheat kind Simeto high-molecular-weight glutelin subunit as shown in Figure 2.
Embodiment 3
1. material sampling
The seed of using club wheat Club20 and bread wheat kind Kontrast respectively is as test material, and the picked at random seed does not have endosperm end 10-15mg and is used for glutelin extraction and Capillary Electrophoresis.And then the sample of making after by mixing in 1: 1 with the high-molecular-weight glutelin subunit that is extracted by the seed of club wheat Club20 and Kontrast respectively is as test material.
2. specimen preparation
High-molecular-weight glutelin subunit (HMW-GS) extracting method:
The standby reagent of selecting for use is identical with embodiment 1 with method of operating, and the HMW-GS that obtains precipitation (w/v:1mg+10 μ l) in proportion adds 20% acetonitrile+0.1%w/v trifluoroacetic acid (TFA), fully dissolving, can carry out capillary electrophoresis analysis after centrifugal; Also precipitation can be put into-20 ℃ of preservations, capillary electrophoresis analysis is carried out in dissolving when need waiting.
3. high performance capillary electrophoresis (HPCE)
Instrument and equipment
The high performance capillary electrophoresis system (be furnished with software and be used for system operation and data processing) that Bio-Rad (Biofocus 3000) company produces.The fused-silica capillary column internal diameter is 50 μ m, and length is respectively 24cm, and skin scribbles the polyimide protective seam, and inwall is coating not all.
Standby reagent
The Capillary Electrophoresis damping fluid
120mm phosphoric acid-glycine buffer (this pH of buffer value is 2.80, is added with 22% acetonitrile and 0.06% hydroxyprolyl-methylcellulose (HPMC)); Distilled water.
The capillary column washing fluid
A.0.1MNaOH
B.1MH
3PO
4
C. distilled water
D. electrophoretic buffer
The capillary douche program
A. new kapillary preprocessor
H
2O 6min
0.1MNaOH 12min
H
2O 6min
1MH
3PO
4 12min
Buffer 75min
B. sample room cleaning procedure
1MH
3PO
4 3min
Buffer 2.5min
C. test cleaning procedure every day
Flushing before the sample introduction:
H
2O 2.5min
0.1MNaOH 6min
H
2O 6min
1MH
3PO
4 6min
Buffer 24min
D. finish the experiment flushing:
1MH
3PO
4 6min
H
2O 6min
0.1MNaOH 2.5min
H
2O 17min
N
2 11min
The Capillary Electrophoresis condition
Voltage: 13.5KV
Temperature: 38 ℃
Sample introduction: 12KV, 6sec
Electrode: by+to-.
Data processing
Utilize Biofocus Integrator software to carry out data processing, simultaneously the electrophoresis result data-switching is become the ASC sign indicating number, carry out the Excel Treatment Analysis.
Club wheat Club20 (2+12) (5+10) separate and identify collection of illustrative plates as shown in Figure 3 for N, 17+18 with bread wheat kind Kontrast for N, 17+18 by the acid capillary electrophoresis of single sample and compound sample (1: 1) HMW-GS.
Claims (3)
1. the acid capillary electrophoresis authentication method of a wheat high-molecular-weight glutelin subunit, comprise material sampling, specimen preparation, high performance capillary electrophoresis and Data Management Analysis, what wherein material sampling, specimen preparation and data processing adopted is conventional method, electrophoretic buffer when it is characterized in that high performance capillary electrophoresis is 100-120mm phosphoric acid-glycine buffer, this pH of buffer is 2.5-2.8, be added with 18-22% nitrile and 0.04-0.06% hydroxyprolyl-methylcellulose, concrete deposition condition parameter is:
Internal diameter capillaceous is 25 or 50 μ m, and length is 24-28cm
Voltage: 10.0-13.5KV
Temperature: 37-40 ℃
Sample introduction: 8-12KV, 6-10sec.
2. the acid capillary electrophoresis authentication method of wheat high-molecular-weight glutelin subunit according to claim 1, it is characterized in that for can the continuous detecting sample and guarantee the accuracy of testing result, when carrying out high performance capillary electrophoresis, kapillary is washed and must operate by cleaning procedure, according to the difference of flushing purpose, cleaning procedure specifically is divided into following four kinds of situations:
(1) will carry out the pre-service flushing before use for new kapillary, cleaning procedure is as follows:
H
2O 4-6min
0.1M?NaOH 9-12min
H
2O 4-6min
1M?H
3PO
4 8-12min
Buffer 50-75min
(2) before the sample introduction kapillary is washed, cleaning procedure is as follows:
H
2O 1.5-2.5min
0.1M?NaOH 4-6min
H
2O 4-6min
1M?H
3PO
4 4-6min
Buffer 16-24min
(3) when once identifying a plurality of sample of mensuration, will wash between twice sample detection, cleaning procedure is as follows:
1M?H
3PO
4 1.5-3min
Buffer 1.5-2.5min
(4) detect the end back kapillary is washed, cleaning procedure is as follows:
1M?H
3PO
4 4-6min
H
2O 4-6min
0.1M?NaOH 1.5-2.5min
H
2O 13-17min
N
2 9-11min
3. the acid capillary electrophoresis authentication method of wheat high-molecular-weight glutelin subunit according to claim 2, it is characterized in that to set up the standard capillary collection of illustrative plates of Wheat HMW-GS, and then form the complete technical system of a cover that wheat high-molecular-weight glutelin subunit is identified by the present invention.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100372570A CN100337113C (en) | 2004-04-30 | 2004-04-30 | Acidic capillary electrophoresis identification method for high molecular weight glutelin subunit of wheat |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100372570A CN100337113C (en) | 2004-04-30 | 2004-04-30 | Acidic capillary electrophoresis identification method for high molecular weight glutelin subunit of wheat |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1570623A true CN1570623A (en) | 2005-01-26 |
| CN100337113C CN100337113C (en) | 2007-09-12 |
Family
ID=34481613
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2004100372570A Expired - Fee Related CN100337113C (en) | 2004-04-30 | 2004-04-30 | Acidic capillary electrophoresis identification method for high molecular weight glutelin subunit of wheat |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100337113C (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1800812B (en) * | 2005-12-28 | 2010-06-02 | 首都师范大学 | Mass spectrum method for identifying low-molecular-weight glutenin subunit of wheat |
| CN102220107A (en) * | 2011-04-19 | 2011-10-19 | 北京鑫诺美迪基因检测技术有限公司 | Sequencer capillary tube cleaning reagent |
| CN101201334B (en) * | 2006-12-15 | 2011-11-16 | 首都师范大学 | Method for identifying wheat variety using wheat water-solubility protein by high-efficiency capillary electrophoresis |
| CN102323320A (en) * | 2011-06-07 | 2012-01-18 | 首都师范大学 | Capillary electrophoresis method for identifying low molecular weight glutelin subunit of wheat and application thereof |
| CN110205398A (en) * | 2019-06-11 | 2019-09-06 | 四川农业大学 | It is a kind of detect wheat high-molecular-weight glutelin Dy10-m619SN subunit KASP labeled primer and application |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5660701A (en) * | 1996-02-29 | 1997-08-26 | Bio-Rad Laboratories, Inc. | Protein separations by capillary electrophoresis using amino acid-containing buffers |
| JP3038184B2 (en) * | 1998-05-27 | 2000-05-08 | 横河アナリティカルシステムズ株式会社 | Method and apparatus for analyzing anions, amino acids and saccharides by capillary electrophoresis |
| CN1247990C (en) * | 1999-12-02 | 2006-03-29 | 海茂株式会社 | Polyacrylamide precast gels for electrophoresis, process for producing the same and electrophoresis method by using the gels |
| CN1207566C (en) * | 2003-01-29 | 2005-06-22 | 天津师范大学 | Method of detecting crop seed purity |
-
2004
- 2004-04-30 CN CNB2004100372570A patent/CN100337113C/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1800812B (en) * | 2005-12-28 | 2010-06-02 | 首都师范大学 | Mass spectrum method for identifying low-molecular-weight glutenin subunit of wheat |
| CN101201334B (en) * | 2006-12-15 | 2011-11-16 | 首都师范大学 | Method for identifying wheat variety using wheat water-solubility protein by high-efficiency capillary electrophoresis |
| CN102220107A (en) * | 2011-04-19 | 2011-10-19 | 北京鑫诺美迪基因检测技术有限公司 | Sequencer capillary tube cleaning reagent |
| CN102323320A (en) * | 2011-06-07 | 2012-01-18 | 首都师范大学 | Capillary electrophoresis method for identifying low molecular weight glutelin subunit of wheat and application thereof |
| CN102323320B (en) * | 2011-06-07 | 2014-01-08 | 首都师范大学 | Capillary electrophoresis method for identifying low molecular weight glutelin subunit of wheat and application thereof |
| CN110205398A (en) * | 2019-06-11 | 2019-09-06 | 四川农业大学 | It is a kind of detect wheat high-molecular-weight glutelin Dy10-m619SN subunit KASP labeled primer and application |
Also Published As
| Publication number | Publication date |
|---|---|
| CN100337113C (en) | 2007-09-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1932041A (en) | The primers specific to cervus elaphus, c. nippon, c. canadensis and rangifer tarandus gene and the method to identify cervi parvum cornu species | |
| CN1944673A (en) | Fluorescent quantitative PCR detecting method for hepatitis B virus and special reagent kit | |
| CN100337113C (en) | Acidic capillary electrophoresis identification method for high molecular weight glutelin subunit of wheat | |
| CN1873019A (en) | Quick method for testing chain reaction of multiple reverse transcription polymerase of lily virus | |
| CN1050021A (en) | The synthetic method of diterpenic lactone | |
| CN1306043C (en) | Reagent and method for detecting leucocythemia susceptibility | |
| CN1824801A (en) | Real-time fluorescent PCR testing primer, probe and immobilized kit for citrus ulcer bacteria and testing process thereof | |
| CN1343516A (en) | Medicine for nourishing liver, kidney and hair and beautifying face and its preparing process | |
| CN1769496A (en) | Method and apparatus for analyzing nucleic acid amplification | |
| CN1191359C (en) | Nucleic acid sequence of high-molecuale glutanin 14 sub-unit gene of wheat and its application | |
| CN100342030C (en) | Method of testing anticancer agent-sensitivity of tumor cells | |
| CN1815199A (en) | Fluid-drop-flowing injection device and quantitative analysing method | |
| CN1800211A (en) | Low molecular weight wheat glutelin subunit, coding gene and use thereof | |
| CN1261762C (en) | Quality determination method for external cultivated bezoar | |
| CN1757739A (en) | Method of expressing heparinase and its special expression carrier | |
| CN1772922A (en) | Method of identifying invasion of south American glim ant and its nucleic acid sequence, probe and reagent kit | |
| CN1296706C (en) | Capillary electrophoresis method for fast identification of wheat varieties | |
| CN1142277C (en) | Method for identifying attenuated chickenpox virous OKA strain or strain originating therein | |
| CN1809638A (en) | Method of detecting beta 3 adrenaline receptor mutant gene and nucleic acid probe and kit therefor | |
| CN1821269A (en) | Wheat seed hardness relative protein and its code gene and application | |
| CN1844408A (en) | Method for screening Down syndrome high risk fetus | |
| CN1557966A (en) | Internal standard gene suitable to exogenous gene detection such as transgene cotton and application thereof | |
| CN100339488C (en) | Detection method of hepatitis B virus genome drug resistance mutation | |
| CN1308446C (en) | Earthworm plasmin multi-component and the separating, purifying and preparing technology thereof | |
| CN1624154A (en) | Process for testing mutating of chondriosome 12s rRNA gene in human tissue microsample by RFLP method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20070912 Termination date: 20130430 |