PCR quick detection method for bee European foul brood
A detection method, the technology of putrefaction disease, applied in the biological field, can solve the problems of time-consuming, judgment easily affected by subjective factors, etc., achieve the effect of simple detection method, control bee disease, and improve the quality of bee products
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[0030] Concrete steps of the present invention are:
[0031] a. Using the CTAB method or the kit method to obtain the European larval rot disease DNA;
[0032] b. Determination of the concentration and purity of European larval rot DNA: take 5 μL of DNA solution and dilute it to 1 mL with double distilled water, and use a nucleic acid protein analyzer or an ultraviolet spectrophotometer to detect the absorbance values A260 and A280 at 260nm and 280nm respectively, so that A260 The ratio to A280 is between 1.7 and 2.0;
[0033] c. Primers:
[0034] EFB1: 5'-CTT TGAACG CCT TAG AGA-3'
[0035] EFB2: 5'-ATC ATC TGT CCC ACC TTA-3'
[0036] EFB3: 5'-TTAACC TCG CGG TCT TGC GTC TCT C-3';
[0037] d. Using the semi-nested PCR amplification method, according to the 16S rRNA gene of Apis coccus hives, design primers EFB1 and EFB2 for the first round of PCR amplification. The fragment size of the obtained PCR theoretical amplification product is 486bp. The amplification result of th...
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