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Method for detecting barley seed purity by two-dimensional electrophoresis technology

A two-dimensional electrophoresis and purity technology, which is applied in the preparation of test samples and electrochemical variables of materials, etc., can solve the problems of malting and beer companies' losses, price differences, and the inability to accurately detect purity, etc., and achieve the effect of accurate detection

Inactive Publication Date: 2014-11-26
DALIAN POLYTECHNIC UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Barley with different purity also has price differences, but the purity cannot be accurately detected only from the appearance and experience, causing malting and brewing companies to easily cause certain losses in the selection of barley raw materials, so the establishment of barley purity detection methods is very necessary

Method used

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  • Method for detecting barley seed purity by two-dimensional electrophoresis technology
  • Method for detecting barley seed purity by two-dimensional electrophoresis technology
  • Method for detecting barley seed purity by two-dimensional electrophoresis technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1. Standard Curve 1 of Barley Purity and Differential Protein 3D Map Height

[0032] The first step, protein extraction:

[0033] Ganpier No.4 purebred barley samples were taken as Ganpier No.4 barley with the purity of 100, 65, 40, 18, and 0% mixed with Shan 2. After manual peeling, grind with liquid nitrogen, take 0.3g sample and add 3 times the volume of extract solution to extract at room temperature for 2h. Centrifuge at 10,000 g for 10 min, take the supernatant and add an equal volume of Tris-balanced phenol at pH 8.8, and extract at room temperature for 30 min. Centrifuge at 10000g for 10min, suck out the supernatant, and extract Tris-balanced phenol again. The two parts of the phenol phase were combined and added with 4 times the volume of cleaning solution A, extracted at -80°C for 3 hours, and centrifuged at 10,000g for 10 minutes. The protein obtained by centrifugation was washed with cleaning solution A and cleaning solution B respectively, centri...

Embodiment 2

[0042] Example 2. Standard Curve 2 of Barley Purity and Differential Protein 3D Map Height

[0043] The first step, protein extraction:

[0044]Ganpier No.4 purebred barley samples were taken as Ganpier No.4 barley with the purity of 100, 65, 40, 18, and 0% mixed with Shan 2. After manual peeling, grind with liquid nitrogen, take 0.3g sample and add 3 times the volume of extract solution to extract at room temperature for 2h. Centrifuge at 10,000 g for 10 min, take the supernatant and add an equal volume of Tris-balanced phenol at pH 8.8, and extract at room temperature for 30 min. Centrifuge at 10000g for 10min, suck out the supernatant, and extract Tris-balanced phenol again. The two parts of the phenol phase were combined and added with 4 times the volume of cleaning solution A, extracted at -80°C for 3 hours, and centrifuged at 10,000 g for 10 minutes. The protein obtained by centrifugation was washed with cleaning solution A and cleaning solution B respectively, centri...

Embodiment 3

[0053] Embodiment three, sample a purity detection

[0054] Take Ganpier No. 4 purebred barley and Dan No. 2 as a mixed species to obtain a Ganpier No. 4 barley sample with a purity of 50%. The extraction and differential protein spot 3D map heights are the same as those in Examples 1 and 2. The protein spot height in area 1 is 1.3, the purity of the area brought into the standard is 49%, and the error is 1%; the height of the protein spot in area 2 is 1.1, the purity of the area brought into the standard is 45.2%, and the error is 4.8%.

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Abstract

The invention provides a method for detecting barley seed purity. The method comprises the steps of: peeling and crushing pure barley and then adding a buffer solution to extract; purifying by a series of steps to obtain soluble protein of barley; and carrying out two-dimensional electrophoresis on different purities of barley which is interbred by manpower, and analyzing a two-dimensional electrophoresis chart to find out the interbred different protein points. The manually interbred three-dimensional (3D) effect picture is analyzed by application of PDQuest software; and the purity of the barley seed can be accurately detected by a standard curve of height and purity curves of different protein points.

Description

technical field [0001] The invention specifically relates to a method for detecting the purity of barley seeds. Background technique [0002] Barley is an important raw material for brewing beer. The purity of the variety is an important indicator of the quality of barley. Barley with different purity may be mixed with other varieties of barley to bring different types of protein, resulting in different brewing characteristics. Therefore, the purity of barley used in beer production has a great influence on the malting process and beer brewing performance. There are also price differences for barley with different purity, but the purity cannot be accurately detected only from the appearance and experience, causing malting and brewing companies to easily cause certain losses in the selection of barley raw materials, so the establishment of a barley purity detection method is very necessary. [0003] In this study, protein two-dimensional electrophoresis technology and PDQues...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/26G01N1/28G01N1/30
Inventor 赵长新刘宝祥尹亚辉安文涛姚继兵张铭振
Owner DALIAN POLYTECHNIC UNIVERSITY
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