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Silver dyeing method

A silver staining and gel technology, applied in the field of silver staining, can solve the problems of cumbersome steps, long time, and large amount of reagents, and achieve the effect of reducing the amount of reagents and improving efficiency

Inactive Publication Date: 2005-03-16
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The purpose of the present invention is to overcome the silver staining method steps of the prior art which are relatively cumbersome, takes a long time, and uses a large amount of reagents, thereby providing a silver staining method with simple steps, high efficiency and fast

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Take out the polyacrylamide gel after electrophoresis, mark the corner of the gel, add it to the aqueous dyeing mixed solution composed of 20v% ethanol, 0.5v% nitric acid, and 0.05wt% silver nitrate for staining for 5 minutes; then use deionized Wash the silver-stained gel with water for 1 minute; wash with 0.1v% formaldehyde, 2wt% Na 2 CO 3 The chromogenic solution of the aqueous solution develops the color until the bands are clear; stop the color development with 10v% acetic acid for 0.5 minutes.

Embodiment 2

[0020] After the polyacrylamide gel after electrophoresis is taken out, the corner of the gel is cut and marked, and added to an aqueous dyeing mixed solution composed of 75v% ethanol, 3v% nitric acid, and 0.8wt% silver nitrate for staining for 10 minutes; then use deionized water Wash the silver-stained gel for 5 minutes; wash with 0.3v% formaldehyde, 4wt% Na 2 CO 3 The chromogenic solution of the aqueous solution develops the color until the bands are clear; stop the color development with 10v% acetic acid for 3 minutes.

Embodiment 3

[0022] After the polyacrylamide gel after electrophoresis is taken out, the corner of the gel is cut and marked, and added to an aqueous dyeing mixed solution composed of 25v% ethanol, 1v% nitric acid, and 0.2wt% silver nitrate for staining for 8 minutes; then use deionized water Wash the silver-stained gel for 3 minutes; wash with 0.2v% formaldehyde, 3wt% Na 2 CO 3 The chromogenic solution of the aqueous solution develops the color until the bands are clear; stop the color development with 10v% acetic acid for 1 minute.

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PUM

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Abstract

The invention relates to a sliver staining method, including: adding gel into staining solution for 5-10 min, the said staining solution containing 20-75v% ethanol,0.5-3v% nitric acid,0.05-0.8wt% silver nitrate, then rinsing silver stained gel for 1-5 min using deionized water, revealing to clear streak using coloring solution, the said coloring solution containing 0.1-0.3v% formaldehyde,2-4wt%Na#-[2]CO#-[3], at last terminating revealing using 10v% acetic acid aqueous solution for 0.5-3 min. The advantages of silver staining method provided by the invention lie in : integrating fixative solution, oxidation solution and revealing solution into one solution thus reducing the reagent volume, simplifying the steps to three. The silver staining time is less than 20 minutes, which promotes the silver staining efficiency, and havs no influence the silver staining sensitivity.

Description

technical field [0001] The invention relates to a silver staining method. technical background [0002] In molecular biology experiments, the establishment of identification methods for polyacrylamide gel electrophoresis results of DNA strives to obtain high sensitivity, and at the same time, the operation is simple and the results are easy to observe. For the staining of DNA bands, ethidium bromide (EB) staining is more commonly used, and the EB on the DNA molecule in the binding channel is excited by ultraviolet radiation to produce fluorescence to achieve the purpose of observation. Although this method is simple and fast, it can meet the needs of most people. The sensitivity required by the experiment, but it has two disadvantages: (1) the carcinogenic effect of EB complicates the manipulation and handling of the gel; Transformation and flinching. [0003] In order to overcome the above disadvantages, a silver staining method with high sensitivity has been developed. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 杨宁曲鲁江吴桂琴李显耀
Owner CHINA AGRI UNIV
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