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Amplification primers and methods for the complete mitochondrial genome sequence of the tick fly

A whole genome, amplification primer technology, applied in the field of genetic engineering, can solve problems such as increased error rate, short product length, time-consuming and labor-intensive, etc., to reduce the probability of amplification errors, reduce amplification workload, and amplify fragments. long effect

Active Publication Date: 2022-07-01
CENT SOUTH UNIV
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Problems solved by technology

The length of the product after the previous primer amplification reaction is relatively short (400-500bp). According to the calculation of the mitochondrial genome size of 15000bp, more than 30 pairs of primers need to be designed to complete the amplification and effective splicing of all fragments, which is time-consuming and laborious. Repeated amplification and sequencing Lead to increased error rate, affecting the splicing results and annotation accuracy after sequencing

Method used

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  • Amplification primers and methods for the complete mitochondrial genome sequence of the tick fly
  • Amplification primers and methods for the complete mitochondrial genome sequence of the tick fly
  • Amplification primers and methods for the complete mitochondrial genome sequence of the tick fly

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Experimental program
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Embodiment

[0033] 1. Materials and methods

[0034] 1.1 Acquisition of experimental materials

[0035] The specimens were collected from Qamdo, Tibet, and the living samples were soaked in absolute ethanol immediately after collection, and stored at -20°C for later use.

[0036] 1.2 DNA extraction

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Abstract

The invention discloses an amplification primer for the whole mitochondrial genome sequence of C. lepidus and an amplification method thereof. The genome of C. lepidus is amplified by PCR through 10 pairs of specific primers shown in SEQ ID NO. 1-20. , the amplified products are subjected to sequencing and sequence splicing after electrophoresis to obtain the complete mitochondrial genome sequence of tick and fly, which can provide a material basis for the prevention and control of wild animals in Sichuan and Tibet, as well as the monitoring of wild animals' health status.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to an amplification primer for the mitochondrial genome sequence of tick fly and an amplification method thereof. Background technique [0002] The tick fly Lipoptena grahami belongs to the order Diptera, the family Hippoboscidae, and the genus Lipoptena. The genus Lepidoptera is a widely distributed obligate blood-sucking insect. Adults parasitize on the surface of large ungulates such as deer, takin, sheep, and horses, and feed on the blood of the host. Because of the infectious bacteria in their bodies, they can transmit ticks and flies. Trypanosomes, Borrelia burgdorferi, bluetongue virus, Bartonella and other pathogens cause severe infection and even death of the host. The lice and flies parasitic on the surface of sheep can also cause the loss of host body weight, wool yield and fur quality, and bring serious economic losses to animal husbandry. Studies have repo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12N15/10C12N15/12
CPCC12N15/11C07K14/43563C12Q1/686C12Q2565/125
Inventor 孟凡明刘子超吴翔蒋立平杨帆
Owner CENT SOUTH UNIV
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