DNA silver staining method in polyacrylamide gel electrophoresis

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...

AI Technical Summary

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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