A rapid identification method of mirror carp
A mirror carp, rapid technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, etc., can solve the problems of pollution and inability to identify the mirror carp genome, etc., to increase feasibility, increase accuracy and reliability, and improve work efficiency Effect
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Embodiment 1
[0030] The gene sequences of mirror carp and other carp species were measured by next-generation sequencing technology. By analyzing the above genome sequencing data and comparing the sequence differences of carp strains, the missing sequence was found, as shown in SEQ NO: 1; the primer design was directly in The upstream and downstream 200bp of the deletion sequence are designed, and the designed primer sequences are shown in SEQ NO:2 and SEQ NO:3.
[0031] The above-mentioned primers provided by the present invention have high efficiency in PCR reaction and strong specificity for the amplification of the missing sequence.
Embodiment 2
[0033] 1) Take the live fish fins of mirror carp, purse red carp, colorful carp, Yellow River carp and Xingguo red carp, set 4 replicates for each fish, use NEB DNA extraction kit to extract the DNA of the fins of the above live fish, the specific operation See the instructions of the kit for steps.
[0034] 2) using the DNA of the above-mentioned mirror carp, purse red carp, colorful carp, Yellow River carp and Xingguo red carp as templates, and carrying out PCR amplification with the following primers (shown in SEQ NO: 2 and SEQ NO: 3),
[0035] Upstream primer 5′-TGCACGTTACTTTTAAACACTCC-3′
[0036] Downstream primer 5′-AAAAAGCAGCATGCCTTACAA-3′
[0037] The primers were designed by the inventors themselves and synthesized by Invitrogen. The PCR reaction system was a 15 μl reaction system, including: 1.5 μl of 10X buffer; 1.5 μl of DNTP mixture (2.5 μm / μl); 2 μl of DNA; 0.5 μl of upstream primer (10 μm concentration) ; downstream primer (concentration 10pm) 0.5μl; sterile w...
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