Ultra-rapid silver dye detecting method of sugarcane molecular marker PAGE gel

Abstract

The invention discloses an ultra-fast silver staining test method of cane molecule tagged PAGE gel, which comprises the following steps: a modified polyacrylamide gel with the concentration of 5 percent is used for the electrophoresis detection of SSR and AFLP tagged PCR amplified primer; the gel after the electrophoresis is placed into a dyeing liquid to be dyed; the gel is taken out from the dyeing liquid, quickly rinsed, and placed into a developing solution to be developed; the developed gel is cleaned by distilled water for two times and dried, and tape reading is implemented; the ultra-fast silver staining test method combines four traditional steps of fixing, dyeing, developing and image fixing into two simplest steps, thus simultaneously reducing rinsing frequency and time, reducing the use of a fixing liquid, an image fixing liquid, acetic acid, absolute ethyl alcohol, sodium carbonate and sodium hyposulfite, not only greatly decreasing silver staining time and reagent cost, but also ensuring high-quality silver staining effect, and compared with other traditional or improved isotopic silver staining methods, the ultra-fast silver staining test method has larger time and cost advantages.

Description

Technical field: [0001] The invention relates to an ultra-fast silver staining detection method for sugarcane molecular marker PAGE gel. Background technique: [0002] The molecular marker technology of sugarcane has developed rapidly in recent years. At present, the most effective molecular marker technology for the research of sugarcane germplasm resources is AFLP and SSR markers. Polyacrylamide gel is often used for the amplified fragments of the two markers. Two methods: silver staining and radioactive imaging. Although radioactive imaging can clearly display the amplified fragments, it is rarely used due to the high cost of the experiment and the relatively great harm to the practitioners; the silver staining method uses formaldehyde to The silver ions are reduced, and the silver ions are combined with the nucleic acid to display the amplification band to achieve the purpose of development, and its resolution can meet the detection requirements. [0003] At present, mo...

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Problems solved by technology

[0003] At present, most of the reported silver staining methods are conventional silver staining methods and improved methods, but there are still many steps, and the reagents used are poisonous with strong alkali and strong acid. good and bad