Molecular marker for identifying amur sturgeon germplasm and application of molecular marker
A molecular marker, sturgeon technology, applied in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc. Effect
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[0034] The preparation method of 1% agar gel of the present invention is as follows:
[0035] Weigh 0.3g of agarose with a balance, pour it into 30ml of 1×TAE buffer, shake well, heat it in a microwave oven until completely dissolved, take it out and let it cool slightly (to prevent EB from volatilizing at high temperature), and add 2 μL of EB. Put the gel plate (Bio-Rad company) in a horizontal position, and then insert the sample comb. Pour the previously prepared agarose gel into the inner groove of the gel plate. Cool for half an hour and take out the comb after the gel is solidified, put the gel into an electrophoresis tank (Bio-Rad Company) and add 1×TAE electrophoresis buffer to submerge the gel. Spot 1 / 5 volume of spotting buffer (bromophenol blue) on the spotting plate.
Embodiment 1
[0037] The method for identifying the germplasm molecular markers of Acipenser schneideri, using Acipenser schnecke, Acipenser dabryi, Acipenser sinensis, Acipenser russiensis, Acipenser siberia, Acipenser sterletii and Acipenser dabryii as research objects, includes the following steps:
[0038] (I) Genomic DNA was extracted from the samples of Sturgeon's sturgeon, Dabry's sturgeon, Chinese sturgeon, Russian sturgeon, Siberian sturgeon, sterlet and Dauric sturgeon, using a marine tissue DNA extraction kit (Tiangen Company).
[0039] The template DNA was then detected by agarose gel electrophoresis:
[0040] Prepare 1% agarose gel, add 2 μL DNA solution to the sample buffer and mix well for electrophoresis. 100V voltage electrophoresis for 25min. After electrophoresis, determine the concentration of the template DNA according to the brightness of the electrophoresis band on the ultraviolet transmission detector, and dilute the template accordingly so that the final concentrat...
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