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DNA barcoding standard detection fragment for identifying vespa mandarinia and application thereof

A standard detection and Vespa golden ring technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, microbial measurement / inspection, etc., to achieve high repeatability, fast speed, and good stability

Inactive Publication Date: 2020-09-11
DALI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no relevant report on the DNA barcoding technology used to identify Vespa aureus in the existing literature.

Method used

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  • DNA barcoding standard detection fragment for identifying vespa mandarinia and application thereof
  • DNA barcoding standard detection fragment for identifying vespa mandarinia and application thereof
  • DNA barcoding standard detection fragment for identifying vespa mandarinia and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 is used to identify the acquisition of the DNA barcode standard gene sequence of Vespa aureus

[0036] 1. Collection, identification and storage of samples to be tested

[0037] The wasp sample to be tested was collected in Dehong City, Yunnan Province. The sample was identified as Vespa mandarinia Smith by morphological methods by Professor Dong Dazhi of Kunming Institute of Zoology, Chinese Academy of Sciences. After the identification was correct, the samples were soaked in absolute ethanol and stored in a -20°C freezer.

[0038] 2. Preparation of experimental instruments and reagents

[0039] 2.1 Experimental instruments and reagents

[0040] HWS12 constant temperature electric heating constant temperature water bath (Shanghai Yiheng Technology Co., Ltd., serial number 131022035);

[0041] SIGMA1-14ED centrifuge (Sigma, Germany, serial number 133828);

[0042] DYCP-31DN electrophoresis instrument (Beijing Liuyi Instrument Factory, batch number 20130816...

Embodiment 2

[0108] Example 2 Identification of samples of different forms and processed products of Vespa aureus

[0109] 1. Collection and storage of the samples to be tested: the pupae and adults of Vespa jinhuanensis purchased in Dehong City, Yunnan Province were taken as the samples to be tested, and part of the adult bodies were quickly dried in an oven to become adult processed products. The adults of Vespa aureus collected from the field in Dehong City, Yunnan Province were used as positive control samples. All samples were identified as Vespa mandarinia Smith by Prof. Dong Dazhi, Kunming Institute of Zoology, Chinese Academy of Sciences according to morphological methods. After the identification was correct, the samples were soaked in absolute ethanol and stored in a -20°C freezer.

[0110] 2. Preparation of experimental instruments and reagents: the same as in Example 1.

[0111] 3. DNA extraction and template preparation: first, pretreat the positive control sample and the sa...

Embodiment 3

[0117] The specificity detection of the DNA bar code molecular identification method of embodiment 3 Vespa aureus

[0118] 1. Collection and storage of samples to be tested

[0119] The samples to be tested were the medicinal materials of Vespa japonica purchased in Dehong City, Yunnan Province, and the medicinal materials of Periplaneta americana purchased in Baoshan City, Yunnan Province. The samples were identified as Vespa mandariniaSmith and Periplaneta americanaLinn. by Prof. Dong Dazhi, Kunming Institute of Zoology, Chinese Academy of Sciences, and Prof. Yang Zizhong, Dali University. The morphologically identified Vespa aureus adults were used as positive control samples. Wipe the sample to be tested with 75% alcohol for disinfection, take a part of the head or chest muscle tissue of the sample to be tested after disinfection, put it in a 1.5mL centrifuge tube, then soak the muscle tissue with absolute ethanol and store it in a -20°C refrigerator.

[0120] 2. Prepara...

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Abstract

The invention discloses a DNA barcoding standard detection fragment for identifying vespa mandarinia and application thereof, and belongs to the field of biological species identification. The invention discloses a discloses a DNA barcoding standard detection gene sequence as shown in SEQ ID NO.1, and a key technology for molecular identification of the insect medicinal material vespa mandarinia by using the CO II gene sequence and a PCR amplification technology. A PCR product is verified through agarose gel electrophoresis and then subjected to sequencing, a sequencing result is subjected tomanual proofreading and sequence splicing, the spliced sequence is compared with the CO II gene sequence disclosed by the invention, and if the homology is 98% or above, the invention can judge that the insect medicinal material to be identified is the vespa mandarinia source. The DNA molecular identification technology provided by the invention realizes rapid, accurate and reliable identificationon fresh or dry medicinal materials, whole worms or part of worms, pupae or larvae or adults, and ensures the safety of the vespa mandarinia as a medicinal material.

Description

technical field [0001] The invention relates to the field of biological species identification, in particular to a DNA barcode standard detection fragment for identifying Vespa aureus and its application. Background technique [0002] Vespa mandarinia Smith is the largest and ferocious predatory carnivorous insect in the family Vespidae of the order Hymenoptera. Bee, Vespa, Vespa, Wasp, etc. Its larvae, whole body and honeycomb can be used as medicine. The name of the medicinal material is bumblebee (whole), bumblebee (larvae), dew hive (hive), and it is mainly used for rheumatic arthralgia. Vespa is sweet and pungent in taste, warm in nature, and mainly treats rheumatic arthralgia. Its adults and larvae have the effect of expelling wind and dampness, and can cure acute and chronic rheumatic pain and rheumatoid arthritis; etc. It is mainly used for pain relief, wind arthralgia, poison treatment, dental venom, breast pain, etc.; while bee venom is mainly used for bee sting ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6869C12N15/11
CPCC12Q1/6888C12Q1/6869C12Q2531/113C12Q2565/125C12Q2563/185
Inventor 杨自忠陈丹许凯歌王音李成功肖怀童建新赵昱
Owner DALI UNIV
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