Test strip kit for detecting duck-derived materials in food and feed and application of test strip kit
A component detection and detection test paper technology, which is applied in the field of test strip kits for duck source component detection, can solve problems such as difficult to distinguish, difficult to implement, and result discrimination obstacles, and achieve high specificity, accurate results, and improved detection sensitivity. Effect
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Embodiment 1
[0020] In an embodiment of the present invention, a test strip kit for detecting duck-derived components in food and feed, including a primer set and a nucleic acid detection test strip with a nucleotide sequence as shown in SEQ ID No.1-2; the primer set The nucleotide sequence is upstream primer 5'-TGTTACAGGAAGTTACTCGCCT-3', as shown in SEQ ID No.1, downstream primer 5'-GCGGAAGATACAAAAAGACACT-3', as shown in SEQ ID No.2; the 5' of this primer set The ends are labeled separately for easy detection, specifically, two different methods of labeling can be selected from biotin molecules, fluorescein isothiocyanate or digoxin. The substrate of the nucleic acid detection test strip is provided with a sample pad, an absorbent pad and a conjugate pad in sequence; the sample pad is composed of a lateral chromatography matrix, a detection strip embedded in the lateral chromatography matrix, and a quality control strip. The chromatographic matrix is nitrocellulose membrane or nylon mem...
Embodiment 2
[0048] The sensitivity of the nucleic acid test strip detection method, the PCR template is 1 / 10, 1 / 10 2 , 1 / 10 3 , 1 / 10 4 , 1 / 10 5 , 1 / 10 6 , 1 / 10 7 , 1 / 10 8 , 1 / 10 9 , 1 / 10 10 After dilution, the PCR system was established for amplification.
[0049] 1 Materials and methods Duck genomic DNA
[0050] 1.2 Primer design
[0051] 1.3 PCR amplification system
[0052] PCR reaction system, including 10×PCR buffer 10 μL, upstream primer 1.8 μL, downstream primer 1.8 μL, HS TaqDNA polymerase 0.5 μL, DNA template 4 μL, ddH 2 O 1.9 μL, a total of 20 μL.
[0053] PCR reaction conditions: pre-denaturation at 95°C for 5 min, 1 cycle; denaturation at 94°C for 30 sec, annealing at 60°C for 1 min, extension at 72°C for 30 sec, amplification for 30 cycles; extension at 72°C for 10 min, storage at 4°C.
[0054] Take 5 μL each for agarose gel electrophoresis. The gel electrophoresis conditions are: 1×TBE buffer, voltage 100V, electrophoresis time 30min. At the same time, take 5 μ...
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