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Primer for authenticating theba impugnata, kit, application of primer and authenticating method

A kit and snail technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problems of lack of scientific identification methods, low identification reliability, etc., and achieve a simple and fast identification method. Effect

Inactive Publication Date: 2017-11-21
李凯兵
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the prior art, there is no scientific identification method for the identification of attacking tea snails, which can only be identified by appearance, and the identification reliability is low

Method used

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  • Primer for authenticating theba impugnata, kit, application of primer and authenticating method
  • Primer for authenticating theba impugnata, kit, application of primer and authenticating method
  • Primer for authenticating theba impugnata, kit, application of primer and authenticating method

Examples

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Embodiment 1

[0033] This embodiment provides a kit for identifying challenged tea snails. The kit includes primers, elution buffer, DNA washing solution and protease. Wherein, the primers include upstream primers and downstream primers, the gene sequence of the upstream primer is shown by SEQ ID NO: 1, the gene sequence of the downstream primer is shown by SEQ ID NO: 2, the gene sequences of the upstream primer and the downstream primer are artificial sequences .

[0034] The kit provided in this example can be used to identify tea snails that attack tea. During identification, primers are used to perform PCR amplification on the DNA of the sample to be detected, and the DNA is sequenced after purification and other steps, and directly compared with the tea attack snails in the prior art. By comparing the gene sequences of the snails or comparing them through the biophylogenetic tree, it can be judged whether the sample to be tested is an attacking tea snail.

Embodiment 2

[0036] This embodiment provides a method for identifying and attacking tea snails, which is carried out with reference to the following steps:

[0037] step one

[0038] Using the biological database NCBI to obtain the corresponding sequences of tea snail-related relative species, the biological software clusterW was used to compare the COI gene fragments of the tea snail attacking tea snails with the COI gene sequence fragments of related relative species. Based on the nucleotide sequence fragment of the mitochondrial COI gene, the first biological phylogenetic tree of the NJ method was constructed using MEGA software.

[0039] step two

[0040] (1) Extract the DNA of the sample to be tested: extract the total DNA of the experimental specimens of the tea snail X and the tea snail Y to be tested respectively (the following DNA extraction processes are all carried out using the E.Z.N.A.MicroElute Genomic DNA Kit kit), extract the tea snail to be tested The method of snail X a...

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Abstract

The invention provides a primer, kit, application and identification method for identifying attacking tea snails, and belongs to the field of biotechnology. The primers include an upstream primer having a nucleotide sequence shown in SEQ ID NO: 1 and an upstream primer having a : The downstream primer of the nucleotide sequence shown in 2. When in use, use the primers to perform PCR amplification on the DNA of the tea snail to be detected, and establish a second biological tree. By comparing with the first biological tree, if the snail to be detected forms a monophyletic group with the known attack tea snail , it can be judged that the tea snail to be detected is an attacking tea snail, and the identification method is quick and easy.

Description

technical field [0001] The invention relates to the field of biotechnology, and in particular to a primer for identifying tea snails attacking them, a kit, application thereof, and an identification method. Background technique [0002] Features of the genus Tea Snail: Shells without navel, solid, spherical or lens-shaped, pale white, most of them have one or more irregular black bands and stripes. The shell faces are threaded and meshed. The mouth of the shell is slightly inclined, in the shape of a half moon, the edge of the mouth is not enlarged, sharp, and the inner side has a ring lip rib. Jaws with 2 or 3 thick ribs. The sagittal capsule is small, the sagittal sac is straight, and the periphery of the cross-section is 4-lobed. The mucous glands are long and tubular, the seminal vesicle has a protrusion, the penis is short and swollen, most of them have flagellates, but occasionally lack them. [0003] Attacking tea snail characteristics: the shell is medium-sized, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888C12Q1/686
Inventor 李凯兵张永宏吴志毅李海林刘海军谈珺马骏胡学难
Owner 李凯兵
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