Xyleborus sp insect gene barcode detection kit and detection method thereof

A detection kit and barcode technology, which is applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve many problems, and meet the needs of simple operation, lower sample quality, and amplification high efficiency effect

Inactive Publication Date: 2017-05-10
LIANYUN PORT IMMIGRATION INSPECTION & QUARANTINE BUREAU PEOPLES REPUBLIC OF CHINA
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  • Xyleborus sp insect gene barcode detection kit and detection method thereof
  • Xyleborus sp insect gene barcode detection kit and detection method thereof

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Embodiment 1

[0043] Embodiment 1, a gene barcode detection kit for the genus Beetle insects, comprising a gene barcode sequence disc and a primer reagent with an amount of more than 10 times The primer sequence is as follows:

[0044] Upstream primer C1-J-2183 (5 , -CAACATTTATTTTTGATTTTTTGG-3 , )

[0045] Downstream primer TL2-N-3014 (5 ,-TCCAATGCACTAATCTGCCATATTA-3 , )

[0046] The gene barcode sequence disk has 37 standard barcode sequences, the sequences of which are shown in sequence 3-sequence 39 in the gene sequence table, and are suitable for the detection of the following 12 kinds of barcode insects:

[0047] Table 1 Names of species

[0048]

[0049] Its matching components also include the following reagents with an amount of more than 10 times: Taqbuffer, MgC1 2 , dNTPmix, TaqDNA polymerase, ddH 2 O, DNA template.

Embodiment 2

[0050] Embodiment 2, method for detecting the barcode of the insect gene of the genus Brutus:

[0051] Seven samples of red wood beetle, bamboo beetle, broad-faced wood beetle, double-spined long beetle, single-cut wood beetle, four-grain beetle, and chestnut-grain beetle were selected as the samples to be tested. Adopt the kit detection of embodiment 1, the steps are as follows:

[0052] (1) Extraction of DNA: The total DNA was extracted by conventional proteinase K digestion. The extraction steps are as follows:

[0053] Put the worm body (place) into an appropriate amount of distilled water, shake it properly, replace the alcohol in the worm body, and then place the worm body on absorbent paper to absorb the water on the worm body surface; put the worm body into a 1.5ml centrifuge Grind the tube quickly with a special small pestle; add 100 μL of non-toxic CTAB separation buffer (PH8.3), grind, then add 100 μL of the above separation solution, and continue grinding until t...

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Abstract

The invention provides a Xyleborus sp insect gene barcode detection kit. The Xyleborus sp insect gene barcode detection kit is used for rapidly detecting 12 kinds of Xyleborus sp, and comprises a primer reagent and a gene barcode sequence disk, wherein 37 standard barcode sequences are saved in the gene barcode sequence disk. The invention further discloses a method for performing Xyleborus sp insect molecular detection method by using the Xyleborus sp insect gene barcode detection kit. By adopting the kit and the method, convenient and efficient Xyleborus sp insect molecular detection can be performed, 12 kinds of Xyleborus sp insects can be distinguished on a molecular level, the use demand of quarantine identification can be met, and high practicability is achieved.

Description

technical field [0001] The invention relates to the technical field of quarantine and identification of Beetle insects, in particular to a gene barcode detection kit for Beetle insects; the invention also relates to a method for detecting the barcode gene of Beetle insects using the kit. Background technique [0002] The genus S. genus (non-Chinese species) is a plant quarantine pest that is imported into China, and foreign S. spp. species have been intercepted by multiple quarantine inspections at ports in my country. It is mainly distributed in subtropical and tropical regions, and is a kind of tree borer pest with important economic significance. Most of the domestic and foreign studies on the wood beetle focus on the morphological classification, and there are few studies on the molecular phylogenetic evolution of this group. [0003] With the development of international trade and tourism and the impact of climate change and other factors, the risk of the spread of har...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6888
Inventor 潘杰邵沛泽颜柳松付茂强魏厚德顾忠盈
Owner LIANYUN PORT IMMIGRATION INSPECTION & QUARANTINE BUREAU PEOPLES REPUBLIC OF CHINA
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