DNA silver staining method

Abstract

The invention discloses a DNA silver staining method. The method comprises the following steps: (1) cutting off power supply after finishing electrophoresis, pouring a buffer solution out from an electrophoresis tank, then taking off a gel plate, putting the rubber plate into a porcelain dish filled with distilled water, and rinsing the gel plate by the distilled water for 2 to 3 times; (2) carrying out silver staining: adding a staining agent to carry out silver staining, shaking ceaselessly, staining for 8 to 12 minutes, and then rinsing by the distilled water for 2 to 3 times; (3) carrying out developing: adding a developing agent to carry out color development, shaking ceaselessly, and developing for 8 to 12 min; (4) rinsing by the distilled water for 2 to 3 times; and (5) carrying out gel shooting: spreading gelatin onto a X-ray film viewer to take a photo. The method integrates the fixing step and the staining step, adopts ethanol to fix, and overcomes the defects that the belt color is lighter when acetic acid is used for fixing, and the acetic acid has penetrating odor, causticity and so on in the prior sliver staining method. Additionally, the method does not need use ethanol and/ or acetic acid to stop developing color, omits the color development stopping step, and solves the problems of penetrating odor and causticity brought by the acetic acid and the like.

Description

technical field [0001] The invention relates to a DNA silver staining method, in particular to a silver staining and color development method for DNA in denaturing polyacrylamide gel (PAGE). Background technique [0002] Polyacrylamide gel electrophoresis (PAGE) is a commonly used method for analyzing protein and nucleic acid samples. Due to its high resolution, it is widely used in the research of molecular biology and related disciplines. Especially in the research of molecular markers such as AFLP, SSR, SNP and the construction of genetic maps. There are many methods for displaying samples after electrophoresis. The detection of target products mainly adopts chromogenic methods. The currently used chromogenic methods are generally as follows: (1) isotope autoradiography, (2) fluorescent dye labeling method, (3) Ethidium bromide staining method. Among them, isotope autoradiography has the advantages of high sensitivity and strong reliability, but it must use steps such as...

AI Technical Summary

Problems solved by technology

Currently, the silver staining methods of Bassam (1991) and Sanguinetti (1994) are more commonly used. At the same time, many researchers have proposed many improved silver staining methods based on their own practice (Rder et al., 1998; Xu Shaobin et al. 2002), mainly Used for polyacrylamide gel electrophoresis staining, also used for agarose gel staining, its sensitivity is 200 times higher than that of EB, but DNA is not suitable for recovery after silver staining

Silver staining is by far the most sensitive staining method. Most of the existing silver staining methods have the following disadvantages: (1) may cause a high background, especially if the water is not pure enough; (2) time-consuming and laborious ( 1~2h); (3) Expensive; (4) More exposure to toxic substances

The whole process is cumbersome and requires the use of various chemical reagents such as deionized water, ethanol, nitric acid, silver nitrate, sodium carbonate, formaldehyde, sodium thiosulfate and acetic acid

The existing silver staining methods all need to use ethanol or / and acetic acid to stop the color development, and many reagents are used, the operation steps are relatively complicated, and the color development time is relatively long

The currently used silver staining method requires fixing solution, developer and staining solution for each dyeing, and the staining solution must be pre-cooled in advance. If the reaction is terminated improperly, as the staining time prolongs, the gel will become darker and darker, reducing the contrast between the DNA band pattern and the background, and the staining effect will be poor.

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