Method for detecting Papaver somniferum L component in food seasoning
A technology for the detection of food condiments and ingredients, which can be used in the determination/inspection of microorganisms, biochemical equipment and methods, etc., and can solve problems such as addiction.
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Embodiment 1
[0026] Detect poppy shells purchased from traditional Chinese medicine stores as follows, using corn flour as the negative control of the experiment:
[0027] Including 1×PCR Master Mix (containing hot-start Taq DNA polymerase, magnesium chloride, dNTP), 400nmol / L upstream and downstream primers, 1.25μL SYBR Green fluorescent dye; upstream primer CODM F: 5'-gctaaactcacgcttgcagaaa-3'; downstream Primer CODM R: 5'-gttacagacgcaccaatattattcaac-3';
[0028] Follow the procedure below for testing:
[0029] (1) Extraction of DNA from the sample to be tested
[0030] A. Weigh 1.00g of the pretreated sample to be tested into a 50mL centrifuge tube; weigh 2.00-4.00g of the semi-solid sample and liquid sample into a 50mL centrifuge tube.
[0031] B. Add 10 mL of CTAB extraction buffer preheated at 65°C and RNase A enzyme (to make the final concentration 10 μg / mL), incubate at 65°C for 30 minutes after inverting and mixing, and invert and mix the centrifuge tube 2 to 3 times during the ...
Embodiment 2
[0046] The artificially added samples of poppy and hot pot seasoning purchased from supermarkets were tested as follows:
[0047] Including 1×PCR Master Mix (containing hot-start Taq DNA polymerase, magnesium chloride, dNTP), 400nmol / L upstream and downstream primers, 1.25μL SYBR Green fluorescent dye; upstream primer CODM F: 5'-gctaaactcacgcttgcagaaa-3'; downstream Primer CODM R: 5'-gttacagacgcaccaatattattcaac-3';
[0048] Follow the procedure below for testing:
[0049] (1) Extraction of DNA from the sample to be tested
[0050] A. Weigh 1.00g of the pretreated sample to be tested into a 50mL centrifuge tube; weigh 2.00-4.00 of the semi-solid sample and liquid sample into a 50mL centrifuge tube.
[0051] B. Add 10 mL of CTAB extraction buffer preheated at 65°C and RNase A enzyme (to make the final concentration 10 μg / mL), incubate at 65°C for 30 minutes after inverting and mixing, and invert and mix the centrifuge tube 2 to 3 times during the period; 12000g Centrifuge for...
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