A kind of dace dna barcode standard detection gene and its application
A standard detection and barcode technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve problems such as lack of data, and achieve the effect of shortening the identification time
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Embodiment 1
[0032]The acquisition of embodiment 1 Chinese carp standard sequence
[0033] 1. Morphological identification and sample collection and preservation of rohu sinensis
[0034] According to the Chinese dace, it has body length, is slightly rod-shaped, and has a high and wide tail peduncle. Snout blunt and protruding, mouth inferior, transversely split. The front part of the upper lip is smooth, covered by the free kiss skin, and there are small papillae on both sides; the inner edge of the free part of the lower lip has many small papillae, the lower lip is separated from the lower jaw, and there is a deep groove in between, and the upper jaw is the upper lip packaged. There is 1 pair of short barbels, and the snout barbels are often degenerated. Dorsal fin iv-10; pectoral fin i-15-17; pelvic fin i-8; anal fin iii-5; dorsal fin front scales 18-24; pericaudal peduncle scales 22-24; lateral line scales 45-47. The back and sides of the body are blue-black, the scales are purple...
Embodiment 2
[0054] 1. Collection, preservation and processing of samples to be tested
[0055] Part of the muscle tissue was cut from the soaked specimen, stored in 95% alcohol and stored in a -20°C refrigerator.
[0056] 2. DNA extraction
[0057] The DNA in the samples was extracted by the phenol-chloroform method and stored at -20°C for future use.
[0058] 3. PCR primer synthesis
[0059] This method selects the sequence of 2 pairs of primers as follows:
[0060] FishF2_t1:5'-TGTAAAACGACGGCCAGTCGACTAATCATAAAGATATCGGCAC-3' (SEQ ID NO.2)
[0061] VF2_t1:5'-TGTAAAACGACGGCCAGTCAACCAACCACAAAGACATTGGCAC-3' (SEQ ID NO.3)
[0062] FishR2_t1:5'-CAGGAAACAGCTATGACACTTCAGGGTGACCGAAGAATCAGAA-3' (SEQ ID NO.4)
[0063] FR1d_t1:5'-CAGGAAACAGCTATGACACCTCAGGGTGTCCGAARAAYCARAA-3' (SEQ ID NO.5)
[0064] 4. PCR reaction system (25ul)
[0065]
[0066]
[0067] 5. PCR reaction procedure
[0068] 95℃*2min,
[0069]
[0070] 72℃*10min
[0071] After the reaction, turn on the PCR instrument...
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