Primers and probes for detecting fox component in food and feed
A fox and probe technology, applied in the field of rapid detection of animal-derived ingredients, can solve problems such as unidentification, discrepancy between the real attributes of the product and the label, and reduction of the amount of raw materials
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Embodiment 1
[0031] Make the real-time fluorescent PCR detection kit of fox component by following formula, and reagent wherein comprises as follows:
[0032] (1) 2×PCR Master Mix
[0033] Including 0.05u / μL Taq DNA polymerase, reaction buffer, 4mmol / L magnesium chloride, 0.4mmol / L dNTP (dATP, dCTP, dGTP, dTTP);
[0034] Wherein the reaction buffer contains tris-hydrochloric acid, 100mmol / L potassium chloride and 2% Triton X-100 of 20mmol / LpH8.8;
[0035] (2) Upstream primer: 10 μmol / L, the sequence is: 5′-TGGAGCATCAGTAGACCTTACAATTT-3′;
[0036] (3) Downstream primer: 10 μmol / L, the sequence is: 5′-GGCGGGAGGTTTTATATTGATAATAG-3′;
[0037] (4) Taqman probe: 10 μmol / L, the sequence is: 5′-CCCTGCACCTGGCCGGAGTC-3′, its 5′ end is labeled with FAM reporter fluorescent group, and its 3′ end is labeled with TAMRA quencher fluorescent group.
[0038] Follow the procedure below for testing:
[0039] (1) Extraction of DNA from the minced meat sample to be tested
[0040] A. Weigh 0.3g minced meat...
Embodiment 2
[0057] Make the real-time fluorescent PCR detection kit of fox component by following formula, and reagent wherein comprises as follows:
[0058] (1) 2×PCR Master Mix
[0059] Including 0.05u / μL Taq DNA polymerase Polymerase, reaction buffer, 4mmol / L magnesium chloride, 0.4mmol / L dNTP (dATP, dCTP, dGTP, dTTP);
[0060] Wherein the reaction buffer contains 20mmol / L tris-hydrochloric acid of pH8.8, 100mmol / L potassium chloride and 2% Triton X-100;
[0061] (2) Upstream primer: 10 μmol / L, the sequence is: 5′-TGGAGCATCAGTAGACCTTACAATTT-3′;
[0062] (3) Downstream primer: 10 μmol / L, the sequence is: 5′-GGCGGGAGGTTTTATATTGATAATAG-3′;
[0063] (4) Taqman probe: 10 μmol / L, the sequence is: 5′-CCCTGCACCTGGCCGGAGTC-3′, its 5′ end is labeled with FAM reporter fluorescent group, and its 3′ end is labeled with TAMRA quencher fluorescent group.
[0064] Follow the procedure below for testing:
[0065] (1) DNA extraction of the feed sample to be tested
[0066] A. Weigh 0.3g feed sample...
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