GB/T 2930.7-2001 Inspection procedures for forage grass seeds and identification of species and varieties
Some standard content:
IC5 65-020-2U
National Standard of the People's Republic of China
GB/T 2930. 1~-2930. 11—2001 Rules for forage seed testing
2001-03-14 Issued
2001-06.01 Implementation
Issued after the National Quality and Technical Supervision
GB/T 293G.72001
This standard is based on the International Seed Testing Association (1NTA) International Seed Testing Machine (19th edition). This standard adopts the International Standard (1STA, 1999) Part VIII: Identification of Plants and Varieties in terms of its main contents. In addition, it includes two species that are not listed in the International Standard and have more important value in China and have suitable inspection methods. In order to make the standard have the advanced scientific nature of international standards, this standard is a series of standards GB/T757C.1~2937.11-2001 forage seed inspection procedures. This series of standards is for the inspection and identification of forage species, and the following parts are composed of the following standards: GB/T 2030. 1—2001
GB/T 2330.2- 2001
GB/T 29:10. 3—2001
GB/T 2930. 42001
GB/T 280. 5—2001
Ga/r 2950. 6- 2:
3/T 2930.7—21
GB/T2959.8-2931
GB/T 2U0.9—26D1
Seed powder test procedure
Grass seed test procedure
Net energy phase
Single seed test procedure
Muzhuo seed test procedure
Muzhong seed test procedure
Teaching common seed test procedures
Reform seed test procedures
Makeup seed test procedures
Teaching seed test procedures
Other particle number determination
Development test
Vigor*Chemical (four physics) pool determination
Health determination
And variety identification
Integration determination
And verification determination
G3/ 296.1:201
GEO—
General Seed Inspection and Testing for Coated Seeds
Testing Notice
The revised standard is different from the international standard. The international standard puts all the test items after the main text, while the part of the standard is in the appendix. The values are written as references for convenience. In this standard, all the consulting items are written as the main content, and the main content is written in the main text. The relevant appendix is written after the main text of the standard. This standard is attached as the appendix of the test standard. The standard is issued by the Ministry of Agriculture of the People's Republic of China. The originating unit of this standard: Inspection and Testing Center of the Ministry of Agriculture and the Institute of Semiconductors of the Ministry of Agriculture (Lanzhou). The main drafter of the standard is Zhu Yan Guo, and the participating persons are Sun Shenghua, Yu Leng, Zi Wen, and Wei Dongxue Jie: 69
GA/2930.7—20C1
ISTA Before
, the biggest risk to the agricultural industry is that the seeds of some products do not meet their production requirements. Seed quality is a combination of different characteristics or states, which are extremely important to seed industry producers, processors, regulators, operators, certification bodies and government agencies responsible for seed management. In all cases, the ultimate goal of inspection is to determine the seed's value for use. Seeds are living products and their condition cannot be accurately predicted like that of living or non-living products. The methods used must be based on scientific knowledge and experience accumulated in seed testing. The required accuracy and reproducibility are due to the date of the verification. This practice defines the standards and helps to determine the quality of products that can be used in international trade. For this reason, the accuracy of the inspection methods used to date has been improved. The accuracy and accuracy of the test results are essential. When the seed trade crosses national borders it is possible that the inspection and testing will be carried out in different countries. Therefore it is very important that all suppliers use a standard method so that reliable inspection results can be obtained within the scope of the approved use. This procedure is divided into two parts: the main body of the regulation and the appendix. The main body of the regulation describes the definitions of the various testing methods and principles adopted by the manufacturer and outlines the methods and procedures to be used. The appendix expands on the definitions and details the procedures and methods set out in the regulation. If the results of the inspection according to this regulation require an international seed inspection certificate from the association, then the production site must comply with the regulation and the interpretation of the provisions of the regulation should be consistent with the relevant details in the appendix. Individual users should use this procedure and its components as much as possible when implementing the national seed quality management program when managing domestic seed trade. Although it is not necessary to use international testing in this case, it is recognised that non-compliance with internationally accepted specifications and conditions may affect the free flow of seed between countries: an ad hoc testing (advance testing) is a requirement for producers of seed for a particular purpose. Such testing may take into account factors such as the species' level, soil type and altitude. For such testing, this Regulation and the Annexes are intended only to provide guidance on specific measures, which are not covered in the relevant literature. The Regulation and Annexes are intended to be used for specific crops. They are not intended to be used for every crop, but are intended to be used for all other crops mentioned. In addition, it is necessary to refer to the equipment of a specific manufacturer for adequate guidance, and this will always be done when ISA considers such equipment to be the best and excludes similar products from other manufacturers. 6
1 Scope
National Standard of the People's Republic of China
Testing Procedure for Forage Seeds
Species and Varieties Identification
Roles for forage seed testing-Verification of speckes and varieties This standard specifies a method for testing the quality of forage seeds, forage seeds and forage crops. GB/T 29307-2001
This standard is applicable to the testing of the quality of forage seeds, forage seeds and forage crops. 2 Referenced Standards
The following standards contain provisions that are referenced in this standard and are therefore the official provisions of this standard. The versions shown are valid at the time of publication of this standard. All standards will be revised. The parties using the following standards shall use the latest version: GE/T 253C.1--20011
Grassroots inspection type
GB/234-2 Example 1 Forage grass species inspection specification B/T817C-1987 Efficacy value
3 Purpose of inspection
To determine whether the sample submitted for inspection is consistent with the species or sum of the species represented and mentioned in the batch. 4 Definitions
This standard adopts the following definitions.
4.1 Authenticity of the sample
Whether the species and variety submitted for inspection are consistent with the requirements of the documents (such as label tubes). 4.2 Variety Vaieyurity
The percentage of seeds of the same variety in the total number of seeds of the plant sample tested. 4.3 Variability
One or more traits (characteristics) are significantly different from the values of the original variety. 5 Principles
The test is valid only when the sender of the sample has a clear description of the variety or cultivar to be tested and there is only one reliable standard sample to be compared with it. The traits for comparison also include morphology, growth, cell culture, etc. When conducting and/or variety identification, seeds can be used, or the seeds of the more mature varieties and standard samples can be taken out of the market in an air-conditioned laboratory, greenhouse, or small-scale cultivation room, and the young plants can be compared with the seedlings of the standard sample of the same adjacent variety under the same environmental conditions during the same period. In special cases, if the results of the determination are indeed reliable (the chromosome doubling effect is determined), it is not necessary to compare with the standard control sample. National Quality and Technical Supervision Bureau 201-C3-14 approved 7n
2001-06-01 clinical
GB/2930.7-2002
When one or more characters of a species or variety are identical (e.g., self-pollinated species), the seeds, seedlings or plants of different species or varieties can be counted. When the species or varieties are not identical (e.g., cross-pollinated species), the obvious variants are counted and a general judgment is made on the authenticity of the test samples. 6 Apparatus and reagents
6.1 Apparatus
The difference between the instruments and methods used in the test varies. 6.1.1 Experimental experiment: Appropriately equip instruments, apparatus and reagents for morphological, physiological and hygroscopic inspection, chemical determination and germination.
6.1.2 Seedling room and culture box: It should be equipped with equipment that can control the environment to induce the development of identification characters of seedlings. 6.1.3 The plots must have the climate, landscape and inflorescence conditions that can enable the cumulative traits to develop normally, and have adequate protection measures against previous diseases:
6-2 Preparation
Typical, potassium iodide, salt, sodium chloride, chlorpyrifos, hydrogen ion, acrylamide, acid electrolysis method. The instruments and reagents required are shown in the Appendix and Appendix (Appendix of standard pile). 7 Inspection procedures
7.1 Inspection site of products
The site of the products to be inspected must comply with the requirements of Table 1. Table 1 Species and cultivars to be tested The samples should be of similar size to those of the genus Pea (Istra), the genus Chrysanthemum (Istra), and other genus Tordaum (Atrma), and other genus Atrium (Bergillus), and other genus B. and other genus species of similar size 7. Species to be tested 7.2-1 The test samples should be taken in accordance with the provisions of 7.2 of G/T 230.1-2C01. At least 4 samples should be taken from the test samples. During the test, duplicates should be observed. Each replicate should not exceed 19 seeds. If the green method is used, The amount of the initial sample can be appropriately reduced. See Appendix A of the standard. 7.2.2. 7.2.2.1 Shape determination. When determining the species and variety based on the seed morphological characteristics, if necessary, it can be assisted by natural selection: When determining the seed color, it can be observed under natural light with a specific spectrum (such as ultraviolet light with a wavelength of 3). The effective identification characteristics are the shape of the grain, the outer part, the color of the grain, the thickness of the membrane groove, the thickness of the nerve, the number of hairs on the axil, the serrations of the dorsal veins, the inner and outer grains, the shape of the grain and the density of the hairs. The grain color of wheat is an effective feature, which can be classified as pure white, yellow, purple, etc. The color of wheat grains can be distinguished under natural light. The main color of wheat grains shows carbon light, and the yellow-white wheat grains do not show fluorescence. GB/T 2930.7—20C1
There are some differences in color and shape between pea and two common beans, which can be used for identification. They can be identified by eye under sunlight or ultraviolet light.
7.2.2.2 Chemical identification
Chemical determination can be used to identify varieties by reacting special chemical reagents with seed-specific components: 7.2.2.2.1 Determination of the biological content of pea and two common beans
The pea and two common beans contain biological reduction, which is a kind of identification symptom. First, put the seeds in water for 24 hours, then cut a thin slice of the seeds and place it on a glass plate to color it, add 1.~% L.2nl of iodine (1% iodine) and 0.6g K-drop. If the rubbing produces brown-red sediment in the water, it means it contains biological magnetism. Remember to count the number of biological organisms:
7.2.2.2.2 Magnetic table HC salt:
Half of the magnetic table can be used for detection and fluorescence. For the seeds that have been harvested under sub-or severe climates, the hydrochloric acid solution can be used to screen the test.
It is said that the human body teaches that (1 part hydrochloric acid and 4 parts hot water are made into a solution) and the seeds are placed on a piece of paper and allowed to air dry for 1 hour. The color of the solution may be sea blue or yellow according to the content. 7.2.2.2-3 The seeds of the big branch (1./) black friend single (1.) and ordinary early purchased raes are stained with the right super seeds inside and outside. The seeds will be determined by the left and right batches and the required seeds. Using the paper method, pre-wet 1824, transfer the seeds to filter paper moistened with 1.5% carbolic acid (5g of water), run the treatment rate group, 25 large branches 11, black grass and ordinary early excitement 2, calibrate and record the color, compare with the standard samples, the bottle color is divided into unrecommended color, Maobang color, grade color, dark grade color, black and black. 7.2.3 The standard reference method of polyacrylamide gel electrophoresis (PAGE) for identifying different varieties of grass is shown in Appendix A (Appendix to the standard). 7.2.4 The standard reference method of polyacrylamide gel electrophoresis (PAGF) for identifying different varieties is shown in Appendix B (Appendix to the standard). 7.3 Pre-test 7.3.1 Laboratory test According to the provisions of B/T 2930.1, at least 433 seeds are taken from the test sample. They are weighed 4 times, with 10 seeds in each batch. The initial determination is carried out using 1 core. When the initial determination cannot produce a conclusion, another 10% of the seeds shall be taken. The seeds are placed on the appropriate growth bed for germination: When the appropriate growth stage is reached, all the samples can be identified. Some need further processing - some can be directly identified. When determining the chromosome multiplex, the samples can be cut and reported or classified according to the color. The seeds are placed on the culture paper with testing paper in the culture plant for development. When the color of the samples reaches the appropriate stage, the color can be identified. The color can be obtained from the color emulsion, and the color can be deepened by mixing sodium ion or aphrodisiac solution, or by irradiating the bacteria with external light! ~21.7.3.2.2 Photoperiod (Loiumr) Most multiflora varieties show fluorescence under UV light, while most perennial varieties do not show any fluorescence. In order to determine the effect of young roots on external filling, the seeds were placed in a suitable non-fluorescent medium in a moist environment at 20°C or 20°C. The seeds were grown in darkness or at light levels not exceeding 0°C. The seeds were arranged separately to prevent the seeds from burning and causing them to show fluorescence. If the traces of fluorescence are not clear, you can use the method of pre-cooling and washing to treat the roots. When the roots have reached a certain degree (the roots of the effective products can be detected!), they can be identified under ultraviolet light. In order to clearly see whether the roots in the paper layer have fluorescence, the roots that do not show fluorescence must be taken out from the filter. Record the number of seedlings showing fluorescence and the number of seedlings without fluorescence and the number of normal seedlings, and fill in the results with the number. 3-2-3 single (eu)
Br 293C.7—2001
purple sheep) and single) are identified by the same method as black family. In the atmosphere containing oxygen, its roots will emit fluorescence. Under ultraviolet light, the roots of black hair grass show yellow-green carbon light, while the roots of black hair grass show blue-green fluorescence. Can it be planted? .2.2 The black hair grass with low-carbon is low in benzene. When the animal is dead, it will be determined by the river. Before identification, 0.1mT of ammonium hydroxide (NHOH) can be collected. Concentrated HH, add heated water and spray the seedlings. 1 1 After that, it is necessary to carry out the next step to ensure that the total weight of the friends is as pure as possible and the color or color of the industry is sold out. 7.4 Temperature or growth room pull sample signing
7.4.1 The ground should be sufficient to ensure that the single plant can grow, and the number of wind ears of the palm or the generation package should be small. The seeds are inoculated with G52!50.1-2, and 2 are harvested. bzxZ.net
7.4.2 Identification
Plant the appropriate container for the seed Ding and keep it in the environment required for the development of the special characters. Under appropriate environmental conditions, when the appropriate development level is recognized, the positive traits of each plant are recorded, and the 7.5-hour plot is pulled to send the designated variety to the inspection pen. Try to sow two single-complex plots in each variety. To avoid loss of harvest, make appropriate cloth strips in different directions and different plots to provide enough planting stockings for accurate identification. The picking method can be adopted. There are enough rows, and the double-row planting method can allow the distribution of the whole operation. It is generally recommended to harvest 15 rows of grass. \, 332m, reduce the error caused by missing and spacing, the narrowing should be adjusted to make the length of the test area equal to the purchase area, and the spacing can be adjusted if necessary. When the single length can be distinguished by special inspection, the seeds can be separated by three or four years of age or temperature rate, and single socks can be obtained. When the seeds have grown to the same size, the spacing can be changed. If the conditions are suitable, the seeds can be directly inserted into the plot on the day of the new year, and then they can be made into single plants through spacing. Check: at least this year's grain socks.The standard items should be used for comparison, and the recovery number must be determined by the number of times and the new statistical place. The plants need to go through the whole growth period to fully develop the phenotype. Therefore, the main process should be extended to the growth period. However, from the beginning of the five-year growth period (not the main forage) or the extraction period (not the main forage), it is the best time to select the sample. During this period, the variation band is tested several times. The hole can be named for another variety or plant! The actual variation is lacking and recorded. If possible, it is best to calculate or account for the plant effect in the plot when the specified system is established. 8 The results are calculated and expressed as
When the specified system is used, the effect rate is 2. The overall effect of the variation found is used to provide a test for the test: If there are more "? 3, then the percentage is rounded to the decimal place: the numerical value is about G/8.7. In the experiment, the results of the test room are not determined by the room, the room or the room. 8.1 Nodule
is the percentage of the seed of the species or variety (of the species) that has been successfully planted (effective) in the trial planting households (planned households), B.2H plot installation
5 is possible, it should be calculated separately! Calculate the success rate of the species, the success rate of the species that has been successfully planted in the trial planting households (planned households), and the success rate of the species that has been successfully planted in the trial planting households (planned households). For the pasture of aligned row planting, when it is difficult to calculate the total number of replacements in some areas, the station Lechuan uses the data from the variety list to express the rapid number of new production
If the trait is measured in the whole operation, it can also overcome the average effect and the value of the overall effect: the variety of false pollen in flowering, the road board can make up for the rapid recovery of the state of the class to accurately determine the variety of the same speed again: the north variety elimination condition ", in the filling of the service I hybrid plant 6 points and the calculation of 3 results, supplement: the real living condition of the group product and comments. In special circumstances, such as when no control sample is set, the stress should be considered A1 principle
GB/T2930./—2001
Appendix 4||t t||(Standard negative recording)
Standard reference method for the separation of protein from single-grain soybean and black grass seeds by propionamide gel electrophoresis (PAGE) according to the standard reference method. The protein of the protein can be separated by light dodecane condensation without any mark. The protein of the protein can be separated by the electrophoresis process. A2 Apparatus and test equipment
A2.1 Receiver
a) Any suitable direct current apparatus (PhrnariGE2/4ial\Preten: b) Centrifuge, centrifuge,
c) Large flat (0.001 g);
l1 Crusher or seal machine:
e) Water-soluble steel
Water-soluble family-style moistening strip, high-temperature drying sample strip
5) Different specifications of kidney, volume selector, test plate, burning forest, penetration tax: burning phase, dropper, culture blood, syringe 25 and needle, etc.,
A2.2 Reagents
All chemical reagents should be of analytical grade or higher: acrylamide (specially purified for electrophoresis) methylene diamine (specially purified for electrophoresis) BISI: dilight Methyl methane (Tre):
hydrochloric acid;
sodium alkyl sulfate
diol;
2-dimethylformamide;
dimethylformamide (DMF);
persulfate (.4FS or riboflavin):
NNNN-toluene Z-amine (TFME):
glacial acid;
tribasic acid (TCA)
PAGE blue-9 or PAGF blue 3A, any material is equivalent to the "Mass Blue R series staining reagents! A2.3.2.1.2.2.2.3.2.1 ... pT1i.K: 30.3g Tris is dissolved in about 2cc of water, and the pH is adjusted to 1.8 with hydrochloric acid. Then add 1mol/L of hydrochloric acid to stop the acid (about 1ml). Then adjust to 250ml and store for 10 minutes. A2.3.35 LIS solution is dissolved in water [when dissolving, it should be handled gently at room temperature], and the volume is adjusted to 1ml. If precipitation occurs, it can be re-dissolved.
42.3.41 Dissolve 1g ammonium monohydrate in 1ml of dense water. This solution must be prepared fresh before the next use. 42.3.5 Sample extraction: Add 20 μL SDS and 12 ng Strong Blue to 12 μg of concentrated gel (A.2.2). Store both samples at room temperature. If precipitation occurs, allow the samples to re-dissolve. A2.3.6 Standard gel fixative: Add 1 mL glacial acetic acid to 10 μL of ethanol and adjust to 1 μL with distilled water. It will take about 20 μL of gel to block the gel. You can use 2% glacial polyoxyethylene, dioxyacetic acid (2.3). A2.3.7 Dense gel stain: Dilute 15% trichloroacetic acid (TCA) to 2.5 mL with water. PAGE blue or a similar reagent should be used as a solvent. 100ml.day)
one piece of film can be stained with more liquid [L]. A3 procedure
a sample of mother is measured 1 grain of god seed, and the mother requires a more accurate estimate of the purity of the variety. If the seed with more wine is determined, the good fruit is only compared with the standard value: if only one of the main components of the seed batch is screened, the seed with less than 1 grain can be determined. For Ding Chen, A3.1 Sample extraction
A3.1.1 Beans
Use an electric crusher (you can also use a ball or purple crusher) to crush the leaves of each variety into fine powder. Prepare the following method for the solution: 17 parts of the original solution (then A2.3.5): 3 1. Add 1 part of ethanol to 140 parts of hot water. Add only enough for the required amount, and add the components to the weighed grade buffer in a ratio of 4g:1.1. Place in a 1.5 m centrifuge to extract the mixture. Lower the chamber temperature to about 1.5 °C. Use a rotating fan to refloat the fine particles. Heat the centrifuge in a water bath. Keep a small key on the lid to prevent the inside from rising. Cool the supernatant and centrifuge for 5 m at 100 °C. Collect the supernatant with ice. 1. ? In the black
, grind 1.5~.0 seeds in a magnetic mill (or a roller grinder or other selective grinder can be used) and then used for analysis. The selected extract can be prepared in the following way: 17 parts slow extraction 15 parts ethanol 10 parts pure basic methyl alcohol 17 parts effective water. Generally, only the amount used on the same day is prepared. The seed powder and the dilution are prepared to be mg; 1.0m: slop. The subsequent treatment method is the same as A3.1.1. 43.7 Preparation
Root electrocoating equipment structure and type installation Dip the clean glue. If the electrosurgical stomach is to be pasted, it is better to be careful in advance, which can make the strips stick more tightly. It is recommended that the thickness of the cavity be 1.5 or thinner: the following preparation method is used to separate the 12.5-degree separation of the 5% daptomycin, 43-2.1 half-proof separation yield
ER/2930.7—7001
drink preparation 4 pieces of 180m>1/(mr×1.nm need to use the following dosage: =F.4mT.ml/.Tris1CH 3,8 line impulse rate (A2.3.1) M:.25mL frontal cavity crushed port (19.6g full soft An-U.26BIS, use quick retention water to make the volume to i:L ten beats of gas volume in the micro-single home gas, and then 3.751.14P (A2..41F.5l. 10KS (A2.3.3 and 7EmLTEME1 white dial with heavy waves), carefully observe the development of the model) after. Glue pleasant amine core, you can also use 25 injection account to discharge glue. Stock liquid port note 3~4 of this part of each piece of dry and policy collection. Inject the upper part of Fengwei separation gel into the world's benefit filling (or wind) and then static 1 (about 1). Inject, and then use the AFS to clean the ASG [i.e. 3.L of ASG.AS group 10mL water
A3.2.2 Concentrated gel
Use water (or 1 liter of propanol!) to absorb the surface of the aldehyde separation gel!.Use a 1:8 ratio of concentrated amine amine to rinse the gel: rinse with water, carefully remove the diluted solution, and then sink it under suction. Prepare 4 microgels using the following amount of liquid: 1/3, pH 6, A2-C1, 267.2mT. Gel microgel (1.0g with 0.57% S added and 67.2m of gas added) in a bottle, then use 3.9m:S (see A.2.4% $1, see 3.3.3 and 50mF ME1 from the original solution). Slide the microgel into the work section in the gel, insert it into the inner tube, make sure that the gas is below, and then let it air dry. Similarly, the high-efficiency APS deep setting [3.2% AI (C.2 APS can swim mf. Naturally, this link can be omitted in the water. For the dry end glue case, it can be expressed as % nuclear element liquid. The effect of the night wave is under the best light. It is necessary to use a light. The amount of auxiliary cutting needs to be determined by the test. The polymerization is required to be completed within m. The control range is 5ml. The standard concentration of the rescue liquid is 7.5mlL nuclear yellow liquid. A3.3 electric ball
electric addition liquid or electric photography grade flushing liquid is 3.014. Effectiveness 1.1SS. Use the filling technique to determine the running rate of 1 work (or plan to slightly heat it up to reduce S. Note that the upper and lower seeds of the electric secretion language are also very protective. Newly prepared
use the comb to pull out the glue (this is quite soft and thin), and then rinse it with a small rinse. Then add the sample to the micro-rinse device (the sample should be added with a small amount of sample size to reduce the cost. The sample size should be determined by the number of samples collected. It is more appropriate to collect more than 5 samples. If the sample is filtered, 5% of the sample can be used as a scan agent in each sample. The buffer can be added to the sample in advance! A3. When the model is used for the station scene rate, at this time, the bottom of the book will be garbage, and the sample will be filled with the medium aid and aid (be careful not to make the sample liquid move). The difficult glue is placed in the hot sample, and the production block is first used with 25 The current of the ice is 15A, and the current moves along the indicator. The indicator moves to the bottom of the charge collection. The temperature should be maintained at 1-20<. The water from the three outlets is cooled and circulated. 43.4 Due to the difference between determination and dyeing, there are several ways to compensate for the difference. It can be used to determine and solidify the white material in the room. If it is not necessary to get it immediately before use, take out the glue from the mold after the ice is fixed or in the fixed concentration (see A2.3. in the well for at least 1h), rinse it with added water for 5iu. Then put it in the color standard (see 42.3. for at least the above money), and use distilled water to determine the color after collection for 23h (preferably with darker colors). TA), and then sealed in a case 7 bag, for inspection or photography, use a sealant, the gel can be kept under ten. If you need to make the dye quickly, you can put it in a high energy gel chamber for a week, and then add acetic acid and 35 2 wing products to dissolve in the water for 30~6iu. 44. Usually, if the method is used, compare the sample and record whether the sample is the same as the standard temperature variety: the product in the registered chamber is matched with a mountain product as a control. The product should have a certain or a certain protein micro-band and a detailed speed, and the reference product should be kept at the same temperature as GB/T 2930. 2001
The film can be used as a qualitative standard. By comparing the standard rubber, the specific characteristics and the uniqueness of the species are known. In addition, the standard protein content in the film is known, and the molecular weight of the main mass spectrum band can be calculated.
Appendix R
(Standard)
Identification of the species of the volatile oil Standard alternative method B1 of PAGE: Principle: The homogenized protein or diol protein from seeds can be separated by polyacrylamide gel electrophoresis (PAGE). The resulting protein band can be used as the "index" of the variety. This method is used to identify the variety. The single seed variety can be tested by the mixing process (i.e., the variety is tested) to determine the purity of the variety. If a very accurate determination of the purity of the variety is required, the larger the sample efficiency. If the instrument is compared with the standard value or only the consistency of one of the main components is used for age verification, then only 5 types of 12 instruments and reagents can be determined. 2.1 Instruments and reagents The actual equipment and identification are related to the equipment. See Appendix A (Standard Time Record) 1: B2.2 Reagents All chemical reagents should be of analytical grade. Preferably, olefins (after special purification by electroplating) are industrially pure acrylamide (after special purification by electroplating) and glacial acid: H Acid:
Anhydrous:
Ascorbic acid;
Overload
Ning G (or Jiao Ning Shi);
Anhydrous:
Ethanol:
Medium alcohol:
Coomassie blue G250 or raBlue G (or similar reagents).B2.3 Solution
B2.3.1 Take the solution
Add a little (or Yi Y) (0.05% W kind: Kang B2.3.2 Electrophoresis solution
Glacial acid 4. A little acid, dilute to 1% with deionized water and store at low temperature. B2.3.3 Spot amine solution
Glacial acid 1ml, ionized water, dilute to 1% and store at low temperature. D2.3.4 Staining solution
GB/T+2930.7-2001
Add Coomassie Brilliant Blue G0 or SerBlue G1p. 18, methanol 250ml and trioxygen acid 1ml 00R volume 80G blood L deionized water
B3 Procedure
B3.1 Sample extraction
Remove the internal and external parts, crush the sample with pliers, and crush the sample into fine particles with a grinder or electric crusher. The fine particles are put into a 1.5m ethylene centrifuge to obtain: add 1U of TLL (see A.2.3.1), mix the fine powder and extract thoroughly, place at room temperature for 2 days or overnight, centrifuge at 14093× for 15 minutes, and take the supernatant for use! 33.2 Gel preparation
The structure and type of the root belt equipment are installed. Clean the liquid membrane and treat the plate with a group first, which can make the active gel pass through the gel easily. The gel plate can be fixed on a plastic support so that it can be kept free during the filling. The method for preparing two gels (1mm×10mm×1.5mm) is as follows: take 6L of the gel filter and add 13.5g of ethylenediamine, 0.1g of ethylenediamine, 6g of propylene glycol, 0.1g of cyclohexane, and 0.005g of iodine. After shaking, add 100ml of gel buffer. For example, add 0.6% (V/V) peroxide (add 0.2ml for every 150ml of gel solution) and quickly pour it into the gel disc. Before adding hydrogen peroxide solution, the gel can be cooled to freezing point: before bonding, draw a sample of acetic acid on the surface of the gel to make the sample form on the upper part of the gel.
The gel lubricating solution should fill the cavity of the gel, or add a layer of gel on the gel to ensure good bonding. The bonding of the gel should be completed within 5-10 minutes.
Take out the sample from the electrophoresis tube, put the sample into the electrophoresis tube, and inject the appropriate amount of baby oil (depending on the instrument used). Add 18L of sample solution to each sample, put the amine mold into the electrophoresis tube to ensure that the sample grid is full of electrodes, and then perform electrophoresis. The required time is: 2CCV (after stabilization) 1min, 50)V (normal) 2Urmn, so that the water circulation of the front part of the sample flows through the cold arm of the electrophoresis tube to maintain the temperature at 15~2℃, 3.4 Fixation and staining
Take out the film from the electrophoresis tube. Disassemble it, and then The staining is completed within 1 day. After an appropriate staining period, the membrane is rinsed in hot water for 2-3 hours to make the stained area clearer, and then photographed. The blue background on the film can be washed off with 1% trichloroethylene. The best method for naming and identifying the alcohol-soluble protein bands is the comparison method, that is, comparing the shape of the protein bands of the sample with the reliable sample before extraction to see if they are consistent. The same sample can be analyzed. There is no unified naming system for alcohol-soluble proteins internationally, and the alcohol-soluble protein bands are counted in sequence or their relative mobility is determined.4% $1 see 3.3.3 and 50mL FME1 self-proclaimed stock solution). Slide the section to the work section in the glue desert, insert the inner well made of him, make sure the comb is below the air, wave, and then let the air contract 1). Similarly, the plan adopts the high-efficiency APS deep setting [3. Prove 2% AI (C.2 APS can swim mf. Naturally, this link can be omitted in the water. For the dry end glue case, it can be expressed as % nuclear element liquid. The effect of the night wave is under the best light. It is necessary to use a light. The amount of auxiliary cutting needs to be determined by the test. The polymerization is required to be completed within m. The control range is 5ml. The standard concentration of the rescue liquid is 7.5mlL nuclear yellow liquid. A3.3 electric ball
electric addition liquid or electric photography grade flushing liquid is 3.014. Effectiveness 1.1SS. Use the filling technique to determine the running rate of 1 work (or plan to slightly heat it up to reduce S. Note that the upper and lower seeds of the electric secretion language are also very protective. Newly prepared
use the comb to pull out the glue (this is quite soft and thin), and then rinse it with a small rinse. Then add the sample to the micro-rinse device (the sample should be added with a small amount of sample size to reduce the cost. The sample size should be determined by the number of samples collected. It is more appropriate to collect more than 5 samples. If the sample is filtered, 5% of the sample can be used as a scan agent in each sample. The buffer can be added to the sample in advance! A3. When the model is used for the station scene rate, at this time, the bottom of the book will be garbage, and the sample will be filled with the medium aid and aid (be careful not to make the sample liquid move). The difficult glue is placed in the hot sample, and the production block is first used with 25 The current of the ice is 15A, and the current moves along the indicator. The indicator moves to the bottom of the charge collection. The temperature should be maintained at 1-20<. The water from the three outlets is cooled and circulated. 43.4 Due to the difference between determination and dyeing, there are several ways to compensate for the difference. It can be used to determine and solidify the white material in the room. If it is not necessary to get it immediately before use, take out the glue from the mold after the ice is fixed or in the fixed concentration (see A2.3. in the well for at least 1h), rinse it with added water for 5iu. Then put it in the color standard (see 42.3. for at least the above money), and use distilled water to determine the color after collection for 23h (preferably with darker colors). TA), and then sealed in a case 7 bag, for inspection or photography, use a sealant, the gel can be kept under ten. If you need to make the dye quickly, you can put it in a high energy gel chamber for a week, and then add acetic acid and 35 2 wing products to dissolve in the water for 30~6iu. 44. Usually, if the method is used, compare the sample and record whether the sample is the same as the standard temperature variety: the product in the registered chamber is matched with a mountain product as a control. The product should have a certain or a certain protein micro-band and a detailed speed, and the reference product should be kept at the same temperature as GB/T 2930. 2001
The film can be used as a qualitative standard. By comparing the standard rubber, the specific characteristics and the uniqueness of the species are known. In addition, the standard protein content in the film is known, and the molecular weight of the main mass spectrum band can be calculated.
Appendix R
(Standard)
Identification of the species of the volatile oil Standard alternative method B1 of PAGE: Principle: The homogenized protein or diol protein from seeds can be separated by polyacrylamide gel electrophoresis (PAGE). The resulting protein band can be used as the "index" of the variety. This method is used to identify the variety. The single seed variety can be tested by the mixing process (i.e., the variety is tested) to determine the purity of the variety. If a very accurate determination of the purity of the variety is required, the larger the sample efficiency. If the instrument is compared with the standard value or only the consistency of one of the main components is used for age verification, then only 5 types of 12 instruments and reagents can be determined. 2.1 Instruments and reagents The actual equipment and identification are related to the equipment. See Appendix A (Standard Time Record) 1: B2.2 Reagents All chemical reagents should be of analytical grade. Preferably, olefins (after special purification by electroplating) are industrially pure acrylamide (after special purification by electroplating) and glacial acid: H Acid:
Anhydrous:
Ascorbic acid;
Overload
Ning G (or Jiao Ning Shi);
Anhydrous:
Ethanol:
Medium alcohol:
Coomassie blue G250 or raBlue G (or similar reagents).B2.3 Solution
B2.3.1 Take the solution
Add a little (or Yi Y) (0.05% W kind: Kang B2.3.2 Electrophoresis solution
Glacial acid 4. A little acid, dilute to 1% with deionized water and store at low temperature. B2.3.3 Spot amine solution
Glacial acid 1ml, ionized water, dilute to 1% and store at low temperature. D2.3.4 Staining solution
GB/T+2930.7-2001
Add Coomassie Brilliant Blue G0 or SerBlue G1p. 18, methanol 250ml and trioxygen acid 1ml 00R volume 80G blood L deionized water
B3 Procedure
B3.1 Sample extraction
Remove the internal and external parts, crush the sample with pliers, and crush the sample into fine particles with a grinder or electric crusher. The fine particles are put into a 1.5m ethylene centrifuge to obtain: add 1U of TLL (see A.2.3.1), mix the fine powder and extract thoroughly, place at room temperature for 2 days or overnight, centrifuge at 14093× for 15 minutes, and take the supernatant for use! 33.2 Gel preparation
The structure and type of the root belt equipment are installed. Clean the liquid membrane and treat the plate with a group first, which can make the active gel pass through the gel easily. The gel plate can be fixed on a plastic support so that it can be kept free during the filling. The method for preparing two gels (1mm×10mm×1.5mm) is as follows: take 6L of the gel filter and add 13.5g of ethylenediamine, 0.1g of ethylenediamine, 6g of propylene glycol, 0.1g of cyclohexane, and 0.005g of iodine. After shaking, add 100ml of gel buffer. For example, add 0.6% (V/V) peroxide (add 0.2ml for every 150ml of gel solution) and quickly pour it into the gel disc. Before adding hydrogen peroxide solution, the gel can be cooled to freezing point: before bonding, draw a sample of acetic acid on the surface of the gel to make the sample form on the upper part of the gel.
The gel lubricating solution should fill the cavity of the gel, or add a layer of gel on the gel to ensure good bonding. The bonding of the gel should be completed within 5-10 minutes.
Take out the sample from the electrophoresis tube, put the sample into the electrophoresis tube, and inject the appropriate amount of baby oil (depending on the instrument used). Add 18L of sample solution to each sample, put the amine mold into the electrophoresis tube to ensure that the sample grid is full of electrodes, and then perform electrophoresis. The required time is: 2CCV (after stabilization) 1min, 50)V (normal) 2Urmn, so that the water circulation of the front part of the sample flows through the cold arm of the electrophoresis tube to maintain the temperature at 15~2℃, 3.4 Fixation and staining
Take out the film from the electrophoresis tube. Disassemble it, and then The staining is completed within 1 day. After an appropriate staining period, the membrane is rinsed in hot water for 2-3 hours to make the stained area clearer, and then photographed. The blue background on the film can be washed off with 1% trichloroethylene. The best method for naming and identifying the alcohol-soluble protein bands is the comparison method, that is, comparing the shape of the protein bands of the sample with the reliable sample before extraction to see if they are consistent. The same sample can be analyzed. There is no unified naming system for alcohol-soluble proteins internationally, and the alcohol-soluble protein bands are counted in sequence or their relative mobility is determined.4% $1 see 3.3.3 and 50mL FME1 self-proclaimed stock solution). Slide the section to the work section in the glue desert, insert the inner well made of him, make sure the comb is below the air, wave, and then let the air contract 1). Similarly, the plan adopts the high-efficiency APS deep setting [3. Prove 2% AI (C.2 APS can swim mf. Naturally, this link can be omitted in the water. For the dry end glue case, it can be expressed as % nuclear element liquid. The effect of the night wave is under the best light. It is necessary to use a light. The amount of auxiliary cutting needs to be determined by the test. The polymerization is required to be completed within m. The control range is 5ml. The standard concentration of the rescue liquid is 7.5mlL nuclear yellow liquid. A3.3 electric ball
electric addition liquid or electric photography grade flushing liquid is 3.014. Effectiveness 1.1SS. Use the filling technique to determine the running rate of 1 work (or plan to slightly heat it up to reduce S. Note that the upper and lower seeds of the electric secretion language are also very protective. Newly prepared
use the comb to pull out the glue (this is quite soft and thin), and then rinse it with a small rinse. Then add the sample to the micro-rinse device (the sample should be added with a small amount of sample size to reduce the cost. The sample size should be determined by the number of samples collected. It is more appropriate to collect more than 5 samples. If the sample is filtered, 5% of the sample can be used as a scan agent in each sample. The buffer can be added to the sample in advance! A3. When the model is used for the station scene rate, at this time, the bottom of the book will be garbage, and the sample will be filled with the medium aid and aid (be careful not to make the sample liquid move). The difficult glue is placed in the hot sample, and the production block is first used with 25 The current of the ice is 15A, and the current moves along the indicator. The indicator moves to the bottom of the charge collection. The temperature should be maintained at 1-20<. The water from the three outlets is cooled and circulated. 43.4 Due to the difference between determination and dyeing, there are several ways to compensate for the difference. It can be used to determine and solidify the white material in the room. If it is not necessary to get it immediately before use, take out the glue from the mold after the ice is fixed or in the fixed concentration (see A2.3. in the well for at least 1h), rinse it with added water for 5iu. Then put it in the color standard (see 42.3. for at least the above money), and use distilled water to determine the color after collection for 23h (preferably with darker colors). TA), and then sealed in a case 7 bag, for inspection or photography, use a sealant, the gel can be kept under ten. If you need to make the dye quickly, you can put it in a high energy gel chamber for a week, and then add acetic acid and 35 2 wing products to dissolve in the water for 30~6iu. 44. Usually, if the method is used, compare the sample and record whether the sample is the same as the standard temperature variety: the product in the registered chamber is matched with a mountain product as a control. The product should have a certain or a certain protein micro-band and a detailed speed, and the reference product should be kept at the same temperature as GB/T 2930. 2001
The film can be used as a qualitative standard. By comparing the standard rubber, the specific characteristics and the uniqueness of the species are known. In addition, the standard protein content in the film is known, and the molecular weight of the main mass spectrum band can be calculated.
Appendix R
(Standard)
Identification of the species of the volatile oil Standard alternative method B1 of PAGE: Principle: The homogenized protein or diol protein from seeds can be separated by polyacrylamide gel electrophoresis (PAGE). The resulting protein band can be used as the "index" of the variety. This method is used to identify the variety. The single seed variety can be tested by the mixing process (i.e., the variety is tested) to determine the purity of the variety. If a very accurate determination of the purity of the variety is required, the larger the sample efficiency. If the instrument is compared with the standard value or only the consistency of one of the main components is used for age verification, then only 5 types of 12 instruments and reagents can be determined. 2.1 Instruments and reagents The actual equipment and identification are related to the equipment. See Appendix A (Standard Time Record) 1: B2.2 Reagents All chemical reagents should be of analytical grade. Preferably, olefins (after special purification by electroplating) are industrially pure acrylamide (after special purification by electroplating) and glacial acid: H Acid:
Anhydrous:
Ascorbic acid;
Overload
Ning G (or Jiao Ning Shi);
Anhydrous:
Ethanol:
Medium alcohol:
Coomassie blue G250 or raBlue G (or similar reagents).B2.3 Solution
B2.3.1 Take the solution
Add a little (or Yi Y) (0.05% W kind: Kang B2.3.2 Electrophoresis solution
Glacial acid 4. A little acid, dilute to 1% with deionized water and store at low temperature. B2.3.3 Spot amine solution
Glacial acid 1ml, ionized water, dilute to 1% and store at low temperature. D2.3.4 Staining solution
GB/T+2930.7-2001
Add Coomassie Brilliant Blue G0 or SerBlue G1p. 18, methanol 250ml and trioxygen acid 1ml 00R volume 80G blood L deionized water
B3 Procedure
B3.1 Sample extraction
Remove the internal and external parts, crush the sample with pliers, and crush the sample into fine particles with a grinder or electric crusher. The fine particles are put into a 1.5m ethylene centrifuge to obtain: add 1U of TLL (see A.2.3.1), mix the fine powder and extract thoroughly, place at room temperature for 2 days or overnight, centrifuge at 14093× for 15 minutes, and take the supernatant for use! 33.2 Gel preparation
The structure and type of the root belt equipment are installed. Clean the liquid membrane and treat the plate with a group first, which can make the active gel pass through the gel easily. The gel plate can be fixed on a plastic support so that it can be kept free during the filling. The method for preparing two gels (1mm×10mm×1.5mm) is as follows: take 6L of the gel filter and add 13.5g of ethylenediamine, 0.1g of ethylenediamine, 6g of propylene glycol, 0.1g of cyclohexane, and 0.005g of iodine. After shaking, add 100ml of gel buffer. For example, add 0.6% (V/V) peroxide (add 0.2ml for every 150ml of gel solution) and quickly pour it into the gel disc. Before adding hydrogen peroxide solution, the gel can be cooled to freezing point: before bonding, draw a sample of acetic acid on the surface of the gel to make the sample form on the upper part of the gel.
The gel lubricating solution should fill the cavity of the gel, or add a layer of gel on the gel to ensure good bonding. The bonding of the gel should be completed within 5-10 minutes.
Take out the sample from the electrophoresis tube, put the sample into the electrophoresis tube, and inject the appropriate amount of baby oil (depending on the instrument used). Add 18L of sample solution to each sample, put the amine mold into the electrophoresis tube to ensure that the sample grid is full of electrodes, and then perform electrophoresis. The required time is: 2CCV (after stabilization) 1min, 50)V (normal) 2Urmn, so that the water circulation of the front part of the sample flows through the cold arm of the electrophoresis tube to maintain the temperature at 15~2℃, 3.4 Fixation and staining
Take out the film from the electrophoresis tube. Disassemble it, and then The staining is completed within 1 day. After an appropriate staining period, the membrane is rinsed in hot water for 2-3 hours to make the stained area clearer, and then photographed. The blue background on the film can be washed off with 1% trichloroethylene. The best method for naming and identifying the alcohol-soluble protein bands is the comparison method, that is, comparing the shape of the protein bands of the sample with the reliable sample before extraction to see if they are consistent. The same sample can be analyzed. There is no unified naming system for alcohol-soluble proteins internationally, and the alcohol-soluble protein bands are counted in sequence or their relative mobility is determined.1SS. Use the filling technique to determine the running rate of 1 (or two times to slightly heat up to reduce S, note that the upper and lower species in the electric secretion language are also very thin and soft, and the rubber worker must be pulled out by the comb (this is quite thin and soft), and then the sample is added to the sample by the micro-irrigation device (the sample is formed into a small rinse, and the sample is added to the sample with a small rinse (the sample is formed into a small rinse, and the ... The sample can be pre-filtered into the buffer solution! A3. When the sample is used at a high rate, the bottom section will be lowered and the sample will be filled with the sample solution (be careful not to stir the sample liquid). The sample is placed in a hot sample and a 25% current is applied to the sample. The current is added by 5A until the sample moves along the indicator. The sample moves to the bottom of the sample. The temperature should be kept at 1-20°C. The sample is cooled by water from the three outlets. The cooling effect is 43.4°C. Several methods related to dyeing can be used to fix and solidify white matter. If it is not necessary to get the color immediately before use, take out the gel from the gel mold or fix it in the concentration (see A2.3.) for at least 1 hour, rinse with diluted water for 5 minutes and then put it in the color standard (see 42.3. for at least 2 hours), then immerse the collected gel in distilled water for 23 hours (preferably darker color), then seal it in a sealed bag for inspection or photography. Use a sealant to keep the gel under normal conditions. If you need to dye quickly, you can put it in a high energy concentration chamber and place it in a cold printing chamber for a week. Then add acetic acid and 35% acetic acid to the product. The sample is usually compared with the standard product. The product should have a certain temperature or micro-band and a certain speed. The reference product should be Y6
GB/T 2930. 2001
The film can be used as a qualitative standard. By comparing the sample with the standard gel, the quality of the whole seed and the distance between the seed and the seed can be determined. In addition, the protein content of the standard gel can be known by the sample. The molecular weight of the main mass spectrum band can be calculated.
Appendix R
(Standard)
Standard method for identification of seed varieties by polyacrylamide gel electrophoresis (PAGE) B1 Principle
The primary protein obtained from the homogenized protein or the diol protein can be separated by polyacrylamide gel electrophoresis (PAGE) and the resulting proteins can be separated. The oral efficacy can be used as the "index effect" of the variety. This supplementary efficacy is used to identify the variety. It can be determined by the new single-grain variety separation process (that is, the variety is tested) to determine the mixed product. If the purity of the variety is required to be determined very accurately, the sample efficacy will be larger. If the instrument is compared with the standard value or only the consistency of one of the main components is used as the age check, then only 5 grains of the variety can be determined. 12 Instruments and reagents
R2.1 Instruments and reagents
The actual equipment used is related to the identification. See Appendix A (Standard Record) 1: B2.2 Reagents
All chemical reagents should be of analytical grade. Preferably, olefins (after special chemical treatment) B2.3 Solution: B2.3.1 Add ethylene glycol solution in a ratio of 75:25 (/), add 1% (or 0.05%) of 1% (or 1% Y) to the solution. Store at low temperature for 1 week or prepare a new one. .2 Electrophoresis solution
4. Add glacial acid, dilute to 1% with deionized water and store at low temperature. B2.3.3 Plasminogen activator solution
Glacial acid, dilute to 1% with deionized water and store at low temperature. D2.3.4 Staining solution
GB/T+2930.7-2001
Add 180ml Coomassie Brilliant Blue G0 or SerBlue G1p., similar to 18, 250ml methanol and 100g trioxide in 80ml deionized water.
B3 Procedure
B3.1 Sample extraction
Remove internal and external parts, crush the sample with pliers, and break the sample into fine pieces with a grinder or electric crusher. Add 1U of TLL to the 1.5m ethylene centrifuge (see A.2.3.1), mix the powder and extract thoroughly, place at room temperature for 2 days or overnight, centrifuge at 14093× for 15 minutes, take the supernatant and use it for electrophoresis, 33.2 Gel preparation
The structure and type of the equipment are installed well. Put the clean liquid membrane, and treat the plate with a group first, which can make the gel electrophoresis easier to separate. The gel disc can be fixed on a plastic bracket, so that it can be kept free during the filling. The method for preparing two gels (1mm×10mm×1.5mm) is as follows: take 6L of the gel filter and add 13.5g of ethylenediamine, 0.1g of ethylenediamine, 6g of propylene glycol, 0.1g of cyclohexane, and 0.005g of iodine. After shaking, add 100ml of gel buffer. For example, add 0.6% (V/V) peroxide (add 0.2ml for every 150ml of gel solution) and quickly pour it into the gel disc. Before adding hydrogen peroxide solution, the gel can be cooled to freezing point: before bonding, draw a sample of acetic acid on the surface of the gel to make the sample form on the upper part of the gel.
The gel lubricating solution should fill the cavity of the gel, or add a layer of gel on the gel to ensure good bonding. The bonding of the gel should be completed within 5-10 minutes.
Take out the sample from the electrophoresis tube, put the sample into the electrophoresis tube, and inject the appropriate amount of baby oil (depending on the instrument used). Add 18L of sample solution to each sample, put the amine mold into the electrophoresis tube to ensure that the sample grid is full of electrodes, and then perform electrophoresis. The required time is: 2CCV (after stabilization) 1min, 50)V (normal) 2Urmn, so that the water circulation of the front part of the sample flows through the cold arm of the electrophoresis tube to maintain the temperature at 15~2℃, 3.4 Fixation and staining
Take out the film from the electrophoresis tube. Disassemble it, and then The staining is completed within 1 day. After an appropriate staining period, the membrane is rinsed in hot water for 2-3 hours to make the stained area clearer, and then photographed. The blue background on the film can be washed off with 1% trichloroethylene. The best method for naming and identifying the alcohol-soluble protein bands is the comparison method, that is, comparing the shape of the protein bands of the sample with the reliable sample before extraction to see if they are consistent. The same sample can be analyzed. There is no unified naming system for alcohol-soluble proteins internationally, and the alcohol-soluble protein bands are counted in sequence or their relative mobility is determined.1SS. Use the filling technique to determine the running rate of 1 (or two times to slightly heat up to reduce S, note that the upper and lower species in the electric secretion language are also very thin and soft, and the rubber worker must be pulled out by the comb (this is quite thin and soft), and then the sample is added to the sample by the micro-irrigation device (the sample is formed into a small rinse, and the sample is added to the sample with a small rinse (the sample is formed into a small rinse, and the ... The sample can be pre-filtered into the buffer solution! A3. When the sample is used at a high rate, the bottom section will be lowered and the sample will be filled with the sample solution (be careful not to stir the sample liquid). The sample is placed in a hot sample and a 25% current is applied to the sample. The current is added by 5A until the sample moves along the indicator. The sample moves to the bottom of the sample. The temperature should be kept at 1-20°C. The sample is cooled by water from the three outlets. The cooling effect is 43.4°C. Several methods related to dyeing can be used to fix and solidify white matter. If it is not necessary to get the color immediately before use, take out the gel from the gel mold or fix it in the concentration (see A2.3.) for at least 1 hour, rinse with diluted water for 5 minutes and then put it in the color standard (see 42.3. for at least 2 hours), then immerse the collected gel in distilled water for 23 hours (preferably darker color), then seal it in a sealed bag for inspection or photography. Use a sealant to keep the gel under normal conditions. If you need to dye quickly, you can put it in a high energy concentration chamber and place it in a cold printing chamber for a week. Then add acetic acid and 35% acetic acid to the product. The sample is usually compared with the standard product. The product should have a certain temperature or micro-band and a certain speed. The reference product should be Y6
GB/T 2930. 2001
The film can be used as a qualitative standard. By comparing the sample with the standard gel, the quality of the whole seed and the distance between the seed and the seed can be determined. In addition, the protein content of the standard gel can be known by the sample. The molecular weight of the main mass spectrum band can be calculated.
Appendix R
(Standard)
Standard method for identification of seed varieties by polyacrylamide gel electrophoresis (PAGE) B1 Principle
The primary protein obtained from the homogenized protein or the diol protein can be separated by polyacrylamide gel electrophoresis (PAGE) and the resulting proteins can be separated. The oral efficacy can be used as the "index effect" of the variety. This supplementary efficacy is used to identify the variety. It can be determined by the new single-grain variety separation process (that is, the variety is tested) to determine the mixed product. If the purity of the variety is required to be determined very accurately, the sample efficacy will be larger. If the instrument is compared with the standard value or only the consistency of one of the main components is used as the age check, then only 5 grains of the variety can be determined. 12 Instruments and reagents
R2.1 Instruments and reagents
The actual equipment used is related to the identification. See Appendix A (Standard Record) 1: B2.2 Reagents
All chemical reagents should be of analytical grade. Preferably, olefins (after special chemical treatment) B2.3 Solution: B2.3.1 Add ethylene glycol solution in a ratio of 75:25 (/), add 1% (or 0.05%) of 1% (or 1% Y) to the solution. Store at low temperature for 1 week or prepare a new one. .2 Electrophoresis solution
4. Add glacial acid, dilute to 1% with deionized water and store at low temperature. B2.3.3 Plasminogen activator solution
Glacial acid, dilute to 1% with deionized water and store at low temperature. D2.3.4 Staining solution
GB/T+2930.7-2001
Add 180ml Coomassie Brilliant Blue G0 or SerBlue G1p., similar to 18, 250ml methanol and 100g trioxide in 80ml deionized water.
B3 Procedure
B3.1 Sample extraction
Remove internal and external parts, crush the sample with pliers, and break the sample into fine pieces with a grinder or electric crusher. Add 1U of TLL to the 1.5m ethylene centrifuge (see A.2.3.1), mix the powder and extract thoroughly, place at room temperature for 2 days or overnight, centrifuge at 14093× for 15 minutes, take the supernatant and use it for electrophoresis, 33.2 Gel preparation
The structure and type of the equipment are installed well. Put the clean liquid membrane, and treat the plate with a group first, which can make the gel electrophoresis easier to separate. The gel disc can be fixed on a plastic b
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