Honey detection composition, kit, detection method and application
The technology of a composition and a kit is applied in the direction of biochemical equipment and methods, and the determination/testing of microorganisms, so as to achieve reliable results, simple operation, and avoid false positive results
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Embodiment 1
[0037] In this example, the specificity and sensitivity of the primers and probes of litchi, longan, acacia, rape, and longan were verified through the following tests.
[0038] Detection of the main reagents:
[0039] Chloroform and isopropanol were respectively purchased from Beijing Liuhetong Company; CTAB lysate (20g / L CTAB, 1.4mol / L NaCl, 0.1mol / L Tris, 0.02mol / L NaCl 2 -EDTA), CTAB precipitation solution (5g / L CTAB, 0.04mol / L NaCl), and 1.2mol / L NaCl were self-prepared in this experiment; Fast Start Universal Probe MasterMix (Rox) was purchased from Roche; primers and probes were purchased from Synthesis by Shanghai Aoke Biotechnology Co., Ltd., etc.
[0040] The main steps of detection:
[0041] 1) DNA extraction
[0042] Test samples: 30 samples, including acacia, litchi, rape, milk vetch, pear, apple, peach, soybean, barley, rice, cucumber, carrot, longan, orange, Vitex, ginkgo, loquat, jujube, wolfberry, rose, Bee, dog, deer, camel, horse, cow, duck, sheep, fish,...
Embodiment 2
[0070] The inventors of the present invention verified the collected honey samples through the following tests.
[0071] Rapeseed honey, litchi honey, acacia honey, longan honey and their mixtures were selected for real-time fluorescent PCR reaction to determine the feasibility, specificity and actual sensitivity of the established real-time fluorescent PCR method.
[0072] In this example, the specific probes used and the experimental operating conditions for real-time fluorescent PCR detection are as described in Example 1.
[0073] Conventionally, when the sample to be tested is amplified, if more than one positive signal occurs three times, it means that the test result is positive.
[0074] Take the mixture of lychee honey and rapeseed honey as an example, mix lychee honey and rapeseed honey in different proportions. Such as Figure 9 As shown, the proportions of litchi honey and rapeseed honey were 100%, 99.9%, 90%, 50%, 10%, 5%, 1% and 0.1%, respectively. Using the N...
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