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Silver staining method of polyacrylamide gel

A technology of polyacrylamide gel and silver staining, applied in the preparation of test samples, measuring devices, instruments, etc., can solve the problems of affecting the progress of the test, long time required, low resolution, etc., and shorten the color development time , save the amount, simplify the effect of silver staining steps

Inactive Publication Date: 2010-01-27
YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the whole process solves the silver staining and developing process after polyacrylamide gel electrophoresis, the operation is simple and convenient, but it takes a long time, the resolution is low, and the large-scale test is time-consuming and laborious, which seriously affects the test results. Progress

Method used

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  • Silver staining method of polyacrylamide gel
  • Silver staining method of polyacrylamide gel
  • Silver staining method of polyacrylamide gel

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Put the above-mentioned gel after electrophoresis into a porcelain plate filled with distilled water, rinse it twice with distilled water, add a staining solution for silver staining, and the staining solution includes 0.2% AgNO3, 1% glacial acetic acid and 10% absolute ethanol , silver staining time is 20min, rinses 2 times with distilled water then, then adds developing solution and develops color, and developing solution comprises 3% anhydrous NaOH and 0.5% formaldehyde, and developing time is 8min, uses digital camera to gel photograph, Such as figure 2 show.

[0030] Implement the second

[0031] Put the above-mentioned gel after electrophoresis into a porcelain plate filled with distilled water, rinse it twice with distilled water, add a staining solution for silver staining, and the staining solution includes 0.2% AgNO3, 1% glacial acetic acid and 10% absolute ethanol , silver staining time is 15min, rinses with distilled water 2 times then, then adds developi...

Embodiment 3

[0033] Put the above-mentioned gel after electrophoresis into a porcelain plate filled with distilled water, rinse it twice with distilled water, add a staining solution for silver staining, and the staining solution includes 0.2% AgNO3, 1% glacial acetic acid and 10% absolute ethanol , silver staining time is 10min, rinses with distilled water 2 times then, then adds developing solution and develops color, and developing solution comprises 3% anhydrous NaOH and 0.5% formaldehyde, and developing time is 8min, uses digital camera to gel photograph, Such as Figure 4 show.

Embodiment 4

[0035] Put the above-mentioned gel after electrophoresis into a porcelain plate filled with distilled water, rinse it twice with distilled water, add a staining solution for silver staining, and the staining solution includes 0.2% AgNO3, 1% glacial acetic acid and 10% absolute ethanol , silver staining time is 5min, rinses 2 times with distilled water then, then adds developing solution and carries out color development, developing solution comprises 3% anhydrous NaOH and 0.5% formaldehyde, and developing time is 8min, uses digital camera to gel photograph, Such as Figure 5 show.

[0036] Silver staining results of different polyacrylamide gel silver staining methods from Figure 1 to Figure 5 It can be seen that the background of silver staining by the conventional silver staining method is shallow, and the target fragments are also blurred and not clear enough, which brings certain difficulties to the accurate judgment of the type. The improved method test used four gradi...

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Abstract

The invention discloses a silver staining method of a polyacrylamide gel, which comprises the following steps of: firstly, putting an electrophoresed gel into a ceramic disc filled with distilled water; secondly, rinsing the gel by using the distilled water for two times; thirdly, adding a staining solution to carry out the solver staining; fourthly, re-rinsing the gel by using the distilled water for two times; fifthly, adding a developing solution to develop; and finally, using a digital camera to take pictures for the gel. By simplifying the silver staining steps, the silver staining method reduces the types of prepared solutions, simplifies a developing process, saves the dosage of a reagent decreasing the cost, and greatly shortens the developing time and improves the dye staining efficiency on the promise of guaranteeing obtaining an ideal target stripe. The obtained target section is clear enough for convenient genetype judgment and is particularly suitable to be used in processes of studying molecular markers and drawing genetic maps.

Description

technical field [0001] The invention relates to a silver staining method of polyacrylamide gel. Background technique [0002] Because polyacrylamide gel electrophoresis uses a small amount of sample, is not easy to diffuse, has high resolution and is easy to observe, it has been widely used in the process of molecular marker research and genetic map construction. The silver staining process after conventional polyacrylamide gel electrophoresis mainly includes the following steps: (1) The gel after electrophoresis is taken out from the glass plate and put into a porcelain dish equipped with distilled water, and rinsed twice with distilled water. (2) Fixing the gel: Add 10% ethanol fixative solution to the dish to submerge the gel, place on a shaker and shake gently for 20 minutes, pour out the fixative solution, and rinse twice with distilled water. (3) Gel oxidation: add 1% nitric acid to oxidize for 5 minutes, recover the nitric acid, rinse twice with distilled water. (4)...

Claims

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Application Information

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IPC IPC(8): G01N1/30G01N27/447
Inventor 罗相忠梁宏伟李忠邹桂伟王长忠
Owner YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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