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Method for identification of barley varieties by two-dimensional electrophoresis technology

A two-dimensional electrophoresis and barley technology, applied in the direction of material analysis, measuring devices, instruments, etc. by electromagnetic means, can solve the problems of difficult identification, difficult application, high identification cost, etc., and achieve the effect of obvious accuracy and stability

Inactive Publication Date: 2013-06-12
DALIAN POLYTECHNIC UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the relatively dense bands, in the case of small differences, it is easy to cause certain difficulties in identification
SSR markers are identified using DNA technology. Although the results are accurate, the requirements for sample DNA extraction techniques and equipment are relatively strict, and the cost of identification is high, so it is difficult to apply in enterprises.

Method used

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  • Method for identification of barley varieties by two-dimensional electrophoresis technology
  • Method for identification of barley varieties by two-dimensional electrophoresis technology
  • Method for identification of barley varieties by two-dimensional electrophoresis technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment one, identification of several barley varieties

[0031] The first step, protein extraction:

[0032] Five kinds of purebred barley (Ganpi No. 4, Ganpi No. 7, Kenmai No. 7, Xiyin No. 1, and Dan No. 2) were manually peeled and ground with liquid nitrogen, and 0.3 g of the sample was added to 0.9 mL of the extract solution at room temperature Extracted for 2h, centrifuged at 10000g for 10min. Take the supernatant and add an equal volume of pH8.8 Tris to balance phenol, extract at room temperature for 30 minutes, and centrifuge at 10000 g for 10 minutes. Add 4 times the volume of cleaning solution A to the phenol phase, extract at -80°C for 3 hours, and centrifuge at 10,000 g for 10 minutes. The protein obtained by centrifugation was washed with cleaning solution A and cleaning solution B respectively, centrifuged three times, and freeze-dried. The dry powder of the obtained whole protein sample was added to 100 μL of lysate, lysed by vortexing at room temper...

Embodiment 2

[0043] Embodiment two, Ganpi No. 6 identification

[0044] The first step, protein extraction:

[0045] The pure-bred barley Ganbe No. 6 was manually peeled and then ground with liquid nitrogen. 0.3 g of the sample was added to 0.9 mL of extract solution for extraction at room temperature for 2 h, and centrifuged at 10,000 g for 10 min. Take the supernatant and add an equal volume of pH8.8 Tris to balance phenol, extract at room temperature for 30 minutes, and centrifuge at 10000 g for 10 minutes. Add 4 times the volume of cleaning solution A to the phenol phase, extract at -80°C for 3 hours, and centrifuge at 10,000 g for 10 minutes. The protein obtained by centrifugation was washed with cleaning solution A and cleaning solution B respectively, centrifuged 3 times, and freeze-dried. The dry powder of the obtained whole protein sample was added to 100 μL of lysate, lysed by vortexing at room temperature for 1 hour, and lysed overnight at 4°C. Centrifuge at 13000ppm at 4°C f...

Embodiment 3

[0053] Example 3, Ganpi No. 4 barley inspection

[0054] The first step, protein extraction:

[0055] The pure-bred barley Ganbeer No. 4 was manually peeled and ground with liquid nitrogen. 0.3 g of the sample was added to 0.9 mL of extract solution for extraction at room temperature for 2 h, and centrifuged at 10,000 g for 10 min. Take the supernatant and add an equal volume of pH8.8 Tris to balance phenol, extract at room temperature for 30 minutes, and centrifuge at 10000 g for 10 minutes. Add 4 times the volume of cleaning solution A to the phenol phase, extract at -80°C for 3 hours, and centrifuge at 10,000 g for 10 minutes. The protein obtained by centrifugation was washed with cleaning solution A and cleaning solution B respectively, centrifuged 3 times, and freeze-dried. The dry powder of the obtained whole protein sample was added to 100 μL of lysate, lysed by vortexing at room temperature for 1 hour, and lysed overnight at 4°C. Centrifuge at 13000ppm at 4°C for 10...

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Abstract

The invention provides a method for identification of barley varieties. The method comprises: peeling and crushing pure barley, then adding a buffer solution to perform extraction, then carrying out purification through a series of steps so as to obtain water-soluble protein of barley, subjecting different varieties of barley protein to electrophoresis by a two-dimensional electrophoresis technology so as to obtain clear and visualized protein maps, and carrying out contrastive analysis to search differential protein of different varieties, thus effectively identifying barley varieties.

Description

technical field [0001] The invention specifically relates to a method for identifying barley varieties. Background technique [0002] Barley is an important raw material for brewing beer, and different barley varieties will have an important impact on malt production and beer quality. The root cause of the differences in barley varieties lies in genetics, so there are general differences in protein levels among different varieties of barley. For different varieties of barley, there are great differences in the malting process and the quality of finished beer. Therefore, the purity of barley varieties used in beer production has a great influence on malting and beer brewing, so malt and beer manufacturers are very urgent to find an accurate and fast method to detect and identify barley varieties. [0003] In the past, the main methods for identifying barley varieties were A-PAGE electrophoresis and SSR markers based on DNA technology, in addition to relying on appearance ex...

Claims

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Application Information

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IPC IPC(8): G01N27/447
Inventor 赵长新刘宝祥安文涛姚继兵尹亚辉张铭振
Owner DALIAN POLYTECHNIC UNIVERSITY
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