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High-efficiency silver staining method for PAGE

A technology of silver staining and formula, applied in the field of molecular genetics, can solve the problems of long time required, cumbersome process, complicated drug configuration, etc., to achieve the effect of shortening processing time and improving experimental efficiency.

Inactive Publication Date: 2015-03-25
INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the traditional silver staining method often requires complicated drug configuration, cumbersome process and long time

Method used

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  • High-efficiency silver staining method for PAGE
  • High-efficiency silver staining method for PAGE
  • High-efficiency silver staining method for PAGE

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Experimental program
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Effect test

Embodiment 1

[0025] 1) Experimental materials

[0026] The experimental materials were Gossypium.hirsutum cultivated species China Cotton Institute 12 and tetraploid wild species Darwin's cotton (G.darwinii) F2 populations, which came from the Cotton Research Institute of Chinese Academy of Agricultural Sciences. SSR primer sequence is swu19357.

[0027] 2) Extraction of cotton genomic DNA

[0028] Soak the cotton seeds in warm water overnight, and then incubate them in a light incubator for about a week until the fresh leaves are unfolded. 0.5g of fresh leaves are selected and quickly milled in liquid nitrogen to a white powder. The extraction method is based on the high content of phenol CTAB method, and slightly improved. The concentration and quality of the extracted DNA were measured by 0.8% agarose gel electrophoresis and UV spectrophotometer. Dilute the DNA stock solution to an appropriate concentration and store it at -20°C for later use.

[0029] 3) PCR amplification reaction and polya...

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Abstract

The invention belongs to the field of molecular genetics and in particular relates to a high-efficiency silver staining method suitable for polyacrylamide gel. The method comprises the following steps: (1) soaking taken gel in stationary liquid, and treating for 2 minutes, wherein the stationary liquid comprises 10 percent Ethanol and 1.0 percent Acetic acid according to a formula; (2) adding the treated gel into 0.2 percent AgNO3 penetrating fluid for treating for 3 minutes, rapidly adding the gel into water, and cleaning for 20 seconds; and (3) adding the gel into a developing solution, and developing within 2 minutes, wherein the developing solution comprises 2 percent NaOH, 20ml of 37 percent HCOH / L and 0.00175 percent Na2S2O3 according to a formula. According to the method disclosed by the invention, the silver staining process can be finished within 10 minutes, the time consumption is almost the lowest in reported silver staining methods, and the experimental efficiency is obviously improved.

Description

Technical field [0001] The invention belongs to the field of molecular genetics, and specifically relates to an efficient silver staining method suitable for polyacrylamide gel. Background technique [0002] Polyacrylamide gel electrophoresis is currently a commonly used method for DNA detection. It has good resolution, can detect even 1bp differences between DNA fragments, and has high throughput. It can be used in large quantities with a small amount of samples. Detection. Silver staining method is a commonly used staining method in polyacrylamide gel electrophoresis due to its advantages of low toxicity and low pollution. [0003] The first application of the staining technique was in the detection of proteins (Switzer et al. 1979), and then the technique was widely used in the detection of nucleic acids (Guo et al. 2008). Nowadays, with the rapid development of research on the genetic location of quantitative traits, the construction of genetic maps, and association analysis,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30G01N27/447
Inventor 崔兴雷孟菲蔡小彦刘方周忠丽王星星王春英王玉红王坤波
Owner INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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