Molecular marker for identifying sphyraena pinguis gunther from sphyraena putnamae and application of molecular marker
A molecular marker, inverted tooth sting technology, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc. Avoid economic losses, high efficiency
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Embodiment 1
[0026] A molecular marker primer and its identification method for distinguishing oily beetle and mandarin beetle:
[0027] 1) The sample populations were selected from the species of the basilisk and the invertebrate collected in the Beibu Gulf, and the DNA of the two sample populations were extracted respectively;
[0028] 2) using the DNA extracted in step 1) as a template, and amplifying through the PCR reaction system with primer F and primer R respectively;
[0029] The composition of described PCR reaction system is:
[0030] Add in order to the 0.2ml centrifuge tube:
[0031]
[0032] The PCR amplification program was pre-denaturation at 94°C for 2 min, (denaturation at 94°C for 15 sec, annealing at 50°C for 15 sec, extension at 72°C for 15 sec) x 38 cycles, extension at 72°C for 3 min, and storage at 4°C for long-term storage.
[0033] 3) Purify and sequence the PCR products, and compare the sequencing results of the two populations to identify their species.
Embodiment 2
[0035] A molecular marker primer and its identification method for distinguishing oily beetle and mandarin beetle:
[0036] 1) The sample populations were selected from the species of the basilisk and the invertebrate collected in the Beibu Gulf, and the DNA of the two sample populations were extracted respectively;
[0037] 2) using the DNA extracted in step 1) as a template, and amplifying through the PCR reaction system with primer F and primer R respectively;
[0038] The composition of described PCR reaction system is:
[0039] Add in order to the 0.2ml centrifuge tube:
[0040]
[0041]
[0042] The PCR amplification program was pre-denaturation at 94°C for 4 min, (denaturation at 94°C for 35 sec, annealing at 53°C for 30 sec, extension at 72°C for 15 sec) x 30 cycles, extension at 72°C for 10 min, and storage at 4°C for long-term storage.
[0043] 3) Purify and sequence the PCR products, and compare the sequencing results of the two populations to identify their...
Embodiment 3
[0045] A molecular marker primer and its identification method for distinguishing oily beetle and mandarin beetle:
[0046] 1) The sample populations were selected from the species of the basilisk and the invertebrate collected in the Beibu Gulf, and the DNA of the two sample populations were extracted respectively;
[0047] 2) using the DNA extracted in step 1) as a template, and amplifying through the PCR reaction system with primer F and primer R respectively;
[0048] The composition of described PCR reaction system is:
[0049] Add in order to the 0.2ml centrifuge tube:
[0050]
[0051] The PCR amplification program was pre-denaturation at 94°C for 5 min, (denaturation at 94°C for 35 sec, annealing at 50°C for 35 sec, extension at 72°C for 35 sec) x 38 cycles, extension at 72°C for 10 min, and storage at 4°C for long-term storage.
[0052] 3) Purify and sequence the PCR products, and compare the sequencing results of the two populations to identify their species.
...
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