DNA silver staining method in polyacrylamide gel electrophoresis

A technology of polyacrylamide gel and polyacrylamide, which is applied in the field of DNA silver staining, can solve the problems of cumbersome process and achieve the effects of improving sensitivity, saving time and reducing cost

Inactive Publication Date: 2010-10-20
NORTHWEST A & F UNIV
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Problems solved by technology

The whole process is cumbersome, using distilled water or deionized water, ethanol, nitric acid, silver nitrate, sodium carbonate, formaldehyde, sodium thiosulfate, acetic acid and other chemical reagents
In recent years, many researchers have explored the process optimization of silver staining and color development of proteins and DNA in PAGE gels, in order to obtain a silver staining method that is simple, fast, sensitive and low in consumption, but the improved method , the whole process of dyeing and color development still needs at least 40-50min

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  • DNA silver staining method in polyacrylamide gel electrophoresis
  • DNA silver staining method in polyacrylamide gel electrophoresis
  • DNA silver staining method in polyacrylamide gel electrophoresis

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Embodiment Construction

[0013] 1, 3 kinds of traditional DNA silver staining methods and the comparison of DNA silver staining method of the present invention

[0014] According to the operation steps of Table 1, 3 kinds of traditional DNA silver staining methods (method 1, method 2 and method 3 in the figure) are compared with the DNA silver staining method of the present invention, and the electrophoresis detection results are as follows figure 1 shown, from figure 1 As can be seen, the dyeing method of the present invention is obviously better than 3 kinds of traditional dyeing methods.

[0015] Detect the microsatellite BM1329 of 400 goats with 8% non-denaturing polyacrylamide gel electrophoresis, use DNA silver staining method staining of the present invention, detection result is as follows figure 2 shown;

[0016] Detect the polymorphism of 400 goats GDF9 gene exon 2 with 12% denatured polyacrylamide gel electrophoresis, use DNA silver staining method staining of the present invention, te...

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Abstract

The invention discloses a DNA silver staining method in polyacrylamide gel (PAGE) electrophoresis, which comprises the steps of soaking polyacrylamide gel with 0.1% AgNO3 for 10-15min, washing with distilled water twice, developing with 0.04% Na2CO3, 2mL of 37% HCOH/L and 2% mixed solution for 5-6min, and flushing twice with distilled water to complete the whole dyeing process. The conventional DNA silver staining method needs 5-6 chemical reagents, prepares 4 solutions and spends 40-60min to complete the dyeing step, but the method only needs 4 chemical reagents, prepares 2 solutions and spends 20min to reach the result the same as that of the conventional dyeing method, and in denaturing polyacrylamide electrophoresis (PAGE), the dyeing method can reduce the amount of DNA to 0.44ng; and in non-denaturing polyacrylamide electrophoresis (PAGE), the amount of DNA can be reduced to 3.5ng. Compared with the conventional dyeing method, the method of the invention saves the time, reduces the cost, and increases the detection sensitivity.

Description

technical field [0001] The invention belongs to an electrophoresis staining method in the field of molecular genetics, and in particular relates to a DNA silver staining method in polyacrylamide gel electrophoresis (PAGE). Background technique [0002] Polyacrylamide gel (PAGE) electrophoresis is a common method for analyzing protein and nucleic acid samples, and it is widely used in basic and clinical research in molecular biology and related disciplines. There are many methods for displaying samples after electrophoresis. Silver staining is widely used because of its high sensitivity, non-toxicity and harmlessness, and the color results can be stored directly with dry gel, especially in recent years in SSCP, DDPCR and In experiments such as DNA sequencing, it shows its superiority. The traditional silver staining method mainly includes the following steps: (1) pretreatment and fixation, (2) staining, (3) color development, (4) termination of the reaction. The whole proce...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30
Inventor 曹斌云姬云涛安小鹏侯金星王建刚宋宇轩
Owner NORTHWEST A & F UNIV
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