Specific primers and methods for distinguishing cotton bollworm and tobacco budworm
A tobacco caterpillar, specific technology, applied in the field of molecular biology, can solve the problems of easy production confusion, interfere with the accuracy of farmland pest prediction and prediction, etc., and achieve the effects of high accuracy, simple operation and wide application.
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Embodiment 1
[0039] Example 1 Identification of the tobacco budworm and the cotton bollworm raised artificially in the room.
[0040] 1) Synthesize the following primers:
[0041] Primer-F: 5'-AACACCAATTAAAACTCCCACAATT-3'
[0042] Primer-R: 5'-TGTGAGCCAGGTTGGTTTCTAT-3'
[0043] 2) Collect 15 larvae of cotton bollworm and tobacco budworm from different families respectively.
[0044] 3) Using Tiangen's Blood / Cell / Tissue Genomic DNA Extraction Kit (Product No.: DP-304-03), extract the genomic DNA of the tested insects according to the kit instructions.
[0045] 4) PCR amplification, the reaction system is as follows:
[0046]
[0047] The reaction program was: pre-denaturation at 95°C for 3 min; denaturation at 94°C for 30 s, annealing at 57°C for 30 s, extension at 72°C for 30 s, and 35 cycles; extension at 72°C for 7 min.
[0048] 5) After the reaction, the PCR reaction tube was taken out, and the PCR reaction product was detected by electrophoresis.
[0049] 6) Sterile water was u...
Embodiment 2
[0053] The cotton bollworm and tobacco budworm of embodiment 2 field collections
[0054] The specific experimental method is as follows:
[0055] 1) Synthesize the following primers:
[0056] Primer-F: 5'-AACACCAATTAAAACTCCCACAATT-3'
[0057] Primer-R: 5'-TGTGAGCCAGGTTGGTTTCTAT-3'
[0058] 2) 15 insects (not known to be cotton bollworm or tobacco budworm) were collected in cotton and pepper fields respectively.
[0059] 3) Using Tiangen's Blood / Cell / Tissue Genomic DNA Extraction Kit (Product No.: DP-304-03), extract the genomic DNA of the test insects according to the kit instructions.
[0060] 4) PCR amplification, the reaction system is as follows:
[0061]
[0062] The reaction program was: pre-denaturation at 95°C for 3 min; denaturation at 94°C for 30 s, annealing at 57°C for 30 s, extension at 72°C for 30 s, and 35 cycles; extension at 72°C for 7 min.
[0063]5) After the reaction, the PCR reaction tube was taken out, and the PCR reaction product was detected by...
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