A primer and kit for early detection of intestinal microsporidia in grouper
A microsporidia, early detection technology, applied in the direction of microorganism-based methods, microorganisms, biochemical equipment and methods, etc., can solve the problems of delayed treatment, loss of the best period of treatment, loss of grouper aquaculture, etc. High detection specificity and sensitivity, and the effect of programming and standardization
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Embodiment 1
[0024] The early detection method of embodiment 1 grouper intestinal microsporidia
[0025] (1) Select about 1 cm of grouper foregut, scrape the intestinal contents, store in 90% absolute alcohol or directly use for the extraction of genomic DNA;
[0026] (2) Use conventional phenol chloroform or commercial fecal sample genome extraction kit to extract genomic DNA from intestinal contents;
[0027] (3) Using the extracted genomic DNA as a template, design and synthesize specific primers for PCR amplification;
[0028] EntF: 5'-GACGTGACTAAAAGAGCGT-3';
[0029] EntR: 5'-AAGTTTCAGTCTTGCGGCT-3'.
[0030] PCR reaction system (25ul):
[0031]
[0032] PCR reaction program:
[0033]
[0034] After the reaction, turn on the PCR instrument, take out the amplified sample, and store it at 4°C.
[0035] (4) Then take an appropriate amount of the PCR amplification product of step 3) and separate it by agarose electrophoresis, observe it under an ultraviolet light after staining ...
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