A kind of dna barcode standard detection gene of catfish plateau loach and its application
A standard detection and barcode technology, which is applied in the direction of recombinant DNA technology, DNA / RNA fragments, microbial measurement / inspection, etc., can solve the problems of no gene sequence and no data related to catfish plateau loach, and achieve the effect of shortening the identification time
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Embodiment 1
[0032] Example 1 Obtaining of the Standard Sequence of Pseudomonas plateau loach
[0033] 1. Morphological identification and sample collection and storage of Pseudomonas plateau loach
[0034]According to the plateau loach, the catfish has a thick body, a broad front end, slightly flattened, a nearly round rear end, and a thin round tail peduncle. Head large, flat, triangular in back view. Mouth large, inferior, arc-shaped. Lips without mastoids, mandible spoon-shaped. Small eyes. The body has no scales, and the skin on the body surface is scattered with short strips and papillary cortical protrusions. The dorsal side of the body is yellowish brown, the abdomen is light yellow, the back and sides of the body have dark brown circles and cloud spots, and all fins have spots and other morphological characteristics, as well as the joint identification of relevant fish taxonomy experts. . Muscle tissue was cut and stored in 95% ethanol in a -20°C refrigerator.
[0035] 2. D...
Embodiment 2
[0051] 1. Collection, preservation and processing of samples to be tested
[0052] Cut part of the muscle tissue of the sample to be tested from the soaked sample, store it in 95% alcohol and store it in a -20°C refrigerator.
[0053] 2. DNA extraction
[0054] The DNA in the samples was extracted by the phenol-chloroform method and stored at -20°C for future use.
[0055] 3. PCR primer synthesis
[0056] This method selects the sequence of 2 pairs of primers as follows:
[0057] FishF2_t1:5'-TGTAAAACGACGGCCAGTCGACTAATCATAAAGATATCGGCAC-3' (SEQ ID NO.2)
[0058] VF2_t1:5'-TGTAAAACGACGGCCAGTCAACCAACCACAAAGACATTGGCAC-3' (SEQ ID NO.3)
[0059] FishR2_t1:5'-CAGGAAACAGCTATGACACTTCAGGGTGACCGAAGAATCAGAA-3' (SEQ ID NO.4)
[0060] FR1d_t1:5'-CAGGAAACAGCTATGACACCTCAGGGTGTCCGAARAAYCARAA-3' (SEQ ID NO.5)
[0061] 4. PCR reaction system (25ul)
[0062]
[0063]
[0064] 5. PCR reaction procedure
[0065]
[0066] After the reaction is over, turn on the PCR instrument, take ...
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