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Method for searching differential expression proteins among different cells or tissues of Rugao yellow chicken based on non-standard quantitative proteomics technology

A technology of differentially expressed proteins and proteomics, applied in the direction of measuring devices, scientific instruments, instruments, etc., can solve the problems of lack of unified and clear description, and achieve the effect of simple and feasible methods, accurate results and strong repeatability

Active Publication Date: 2020-09-11
YANGZHOU UNIV
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AI Technical Summary

Benefits of technology

In this new way we have developed an improved technique that allows us to find specific types of molecules called certain genes within each cell/tissue found inside red turkeys (Rho). By comparing these genetic material from various parts of their body taken together they will be able to identify differences therebetween - even if it's just one part per million across all three organs being studied at once. These techniques make research faster than current methods like Western blotting but still work well enough when studying small samples such as young birds.

Problems solved by technology

This patents discusses how proteomic techniques have developed over recent years that help scientists better understanding complex systems like living organisms. These methods involve extracting specific parts from their body fluids (blood) and performing various measurements to determine important properties related thereto. By comparing these data sets against each other, they provide valuable insights regarding genetics and biochemistry processes involved in health care applications.

Method used

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  • Method for searching differential expression proteins among different cells or tissues of Rugao yellow chicken based on non-standard quantitative proteomics technology
  • Method for searching differential expression proteins among different cells or tissues of Rugao yellow chicken based on non-standard quantitative proteomics technology
  • Method for searching differential expression proteins among different cells or tissues of Rugao yellow chicken based on non-standard quantitative proteomics technology

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Embodiment Construction

[0028] (1) In this example, Rugao yellow chicken ESCs and PGCs cells were selected to be isolated. After separation, they were quickly placed in liquid nitrogen for quick freezing. The quick-frozen samples were stored in a -80°C refrigerator. After all samples were collected, the samples were collected from Take it out at -80°C, add 4 times the volume of lysis buffer (8M urea, 1% protease inhibitor, 50uM PR-619), and ultrasonically lyse. Centrifuge at 12,000 g for 10 min at 4°C to remove cell debris, transfer the supernatant to a new centrifuge tube, and use the BCA kit for protein concentration determination.

[0029] (2) Add dithiothreitol to the protein solution to make the final concentration 5mM, and reduce at 56°C for 30min. Afterwards, iodoacetamide was added to make the final concentration 11 mM, and incubated at room temperature in the dark for 15 min. Finally the urea concentration of the sample was diluted below 2M. Add trypsin at a mass ratio of 1:50 (trypsin:pro...

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Abstract

The invention relates to a method for searching differential expression proteins among different cells or tissues of Rugao yellow chicken based on a non-standard quantitative proteomics technology. The method comprises the following steps: (1) extracting tissues or cell proteins of different cells or tissues of chicken; (2) carrying out pancreatin enzymolysis on different groups of cells or tissueproteins obtained in the step (1); (3) carrying out liquid chromatography-mass spectrometry analysis on the cell or tissue proteins subjected to enzymolysis; and (4) performing database search and annotation analysis on an analysis result. According to the present invention, with the method, the differential expression proteins between different tissues and cells can be rapidly obtained; 2, the method is simple, feasible, high in repeatability and accurate in result; and 3, the method is suitable for other species.

Description

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Claims

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Application Information

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Owner YANGZHOU UNIV
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