Molecular marker method for cerambycidae larvae of different families

A molecular marker and pedigree technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of lack of larvae isolation and identification data, confusing identification, and undescribed larval morphology.

Inactive Publication Date: 2016-03-30
太仓海关综合技术服务中心
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Problems solved by technology

[0002] The taxonomic research of longhorn beetle larvae mainly focuses on the field of individual morphology, but due to the extreme lack of larvae isolation and identification data, many species of longhorn beetle larvae have not been described so far; the same species of longhorn beetle larvae with different geographical distributions have certain differences in morphology; Close species, very similar in shape, easy to be confused when identifying

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  • Molecular marker method for cerambycidae larvae of different families
  • Molecular marker method for cerambycidae larvae of different families
  • Molecular marker method for cerambycidae larvae of different families

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Embodiment 1

[0026] 1. Test material:

[0027] The longicorn beetle specimens used were the specimens intercepted by the insect laboratories of Jiangsu Entry-Exit Inspection and Quarantine Bureau for many years. The sources and collection times of the five specimens are shown in Table 1. In addition, 28 beetle COI gene sequences registered in Genbank were selected for comparison, see Table 2.

[0028] Table 1 Name and source of experimental specimens

[0029]

[0030] Table 2 Genbank download sequence information

[0031]

[0032]

[0033] 2. Extraction of total genomic DNA

[0034] Using GenMag Animal Tissue / Cell Genomic DNA Extraction Kit: soak the worm body in alcohol, select muscle tissue of appropriate size, wash 2-3 times with sterile water, drain the water, grind the tissue, add appropriate amount of lysate Solution and proteinase K, 55 ℃ warm bath, so that the tissue is completely lysed. Then add magnetic beads and buffer to make the DNA adsorb to the magnetic beads, ...

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Abstract

The invention discloses a molecular marker method for cerambycidae larvae of different families. The method is realized by the steps of: selecting two pairs of specific primers to carry out PCR reaction, and extracting genomic DNA from to-be-detected cerambycidae individuals and parents of different families by DNA extraction; taking the genomic DNA as the template, and carrying out two rounds of PCR amplification on the selected two pairs of primers respectively; subjecting the PCR product to 1.5% agarose electrophoresis detection, performing ethidium bromide staining, then conducting observation under ultraviolet, and photographing the electrophoresis result; and constructing a phylogenetic tree by a neighbor-joining method. According to the method provided by the invention, the genetic distances of 33 cerambycidae COI genes indicate that: the genetic distance between different genuses is obviously larger than that of a same genus, the COI gene is in full compliance with the inspection standard of DNA bar code validity, and the fragment can well distinguish different species of cerambycidae's different genuses, and can be used for rapid identification of cerambycidae.

Description

technical field [0001] The invention belongs to the technical field of molecular markers, and in particular relates to a molecular marker method for different families of cervix larvae. Background technique [0002] The taxonomic research of longhorn beetle larvae mainly focuses on the field of individual morphology, but due to the extreme lack of larvae isolation and identification data, many species of longhorn beetle larvae have not been described so far; the same species of longhorn beetle larvae with different geographical distributions have certain differences in morphology; Close species, very similar in shape, easy to confuse identification. Therefore, along with the development of molecular biology, nucleic acid sequence analysis (DNA sequence analysis) is used more and more in the molecular identification of longhorn beetle larvae, which is of great significance in entry-exit inspection and quarantine work. [0003] Mitochondrial DNA (mtDNA) has a small molecular ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 顾忠盈吕飞
Owner 太仓海关综合技术服务中心
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