Preparation method of polyacrylamide gel for molecular marker identification

Abstract

The invention discloses a preparation method of polyacrylamide gel for molecular marker identification. The preparation method comprises the following steps: 1) preparing and storing 26% polyacrylamide mother liquor; 2) preparing and storing 1.5M Tris-HCl (having a pH value of 8.8); 3) preparing and storing 10%g / ml APS; 4) preparing and storing a polyacrylamide gel working solution; 5) adding 10%APS and TEMED (Tetramethylethylenediamine) into the polyacrylamide gel working solution before use so as to obtain a mixture, wherein a ratio of the polyacrylamide gel working solution to the 10% APSto the TEMED is 30ml:200uL:50uL; and 6) performing electrophoresis. The preparation method disclosed by the invention is simple in process and high in efficiency, the polyacrylamide gel has high stability, the working solution can be stored for a long time, the pharmaceutical cost is greatly saved, the gel preparation efficiency is improved, the simplicity and stability of molecular marker identification are greatly ensured, and a guarantee is provided for polymorphism identification of molecular markers. Meanwhile, by utilizing the preparation method disclosed by the invention, the experimental precision and accuracy can be improved, and the experimental cost and workloads of experimenters are reduced.

Description

Technical field [0001] The invention relates to the technical field of molecular marker identification, in particular to a simple, stable and storage-resistant polyacrylamide gel preparation method for molecular marker identification. Background technique [0002] The identification of molecular markers is the most convenient and commonly used method in molecular marker-assisted selection, variety purity screening, and gene cloning. At present, commonly used molecular markers are AFLP, RFLP, SSR, SNP, and Indel. Among the above-mentioned molecular markers, except for SNP molecular markers that can be identified by a fluorescent quantitative PCR instrument based on the melting curve, the identification of other molecular markers almost all rely on polypropylene gel electrophoresis. In actual experiments in the prior art, due to the high cost of reagents and consumables required for the identification of SNPs by melting curves, the identification of SNP molecular markers using CAP...