Propylaea Japonica specific COI primers, kit containing primer and detection method thereof

A mole ladybird and detection method technology, applied in the field of molecular biology, can solve the problems of impact, population dynamics, complex biological control effects of pests, inability to accurately evaluate group predation among ladybugs, etc., and achieve simple operation and high practical value , high accuracy effect

Inactive Publication Date: 2013-12-18
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Application Information

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Problems solved by technology

[0003] With the in-depth research on the predation behavior of ladybugs, it is found that the phenomenon of intra-group predation is common among ladybugs, and this phenomenon has a complex and huge impact on the population dynamics of different species in farmland ecosystems and the effect of biological control of pests
Due to the wide variety of ladybugs, traditional research methods such as rout

Method used

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  • Propylaea Japonica specific COI primers, kit containing primer and detection method thereof
  • Propylaea Japonica specific COI primers, kit containing primer and detection method thereof
  • Propylaea Japonica specific COI primers, kit containing primer and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] The amplification effect of embodiment 1 primer PjF1 / PjR1 to the moire ladybug

[0024] (1) Preparation of the mole beetle genome

[0025] Put the single-headed ladybug into a 1.5mL centrifuge tube and grind it, and use TIANamp Genomic DNA Kit Blood / Cell / Tissue Genomic DNA Extraction Kit (spin column type) (Tiangen Biological Co., Ltd., Beijing) to extract single The head ladybug genome. The DNA solution was stored at -20°C for later use, and 1 μL of the solution was taken as a DNA template during PCR amplification.

[0026] (2) Synthesis of specific COI primers for the detection of mole beetle

[0027] The specific COI primer sequences are as follows:

[0028] PjF1: 5'-AAATGACCAAATTTATAATGTT-3' (SEQ ID No. 1)

[0029] PjR1: 5'-TGATGATAAAGGAGGATAAACT-3' (SEQ ID No. 2)

[0030] (3) PCR amplification

[0031] The reaction system is 20 μL, including: 10×EasyTaq buffer 2.0 μL, dNTP (2.5 mM) 0.4 μL, EasyTaq DNA polymerase (5U / μL) 0.2 μL, forward and reverse primers (10...

Embodiment 2

[0040] Example 2 Primer PjF1 / PjR1 Determination of the Minimum Detection Amount of Coccinella moriensis

[0041] 1) Preparation of the template DNA of the mole-shaped ladybug

[0042] According to Example 1, the single-headed ladybug genomic DNA was extracted, and then the original template solution (the concentration of the DNA solution was 3ng / μL) was gradually diluted by 2 times until no bands were detected, and 1 μL was used as a template for PCR amplification. Add directly to the PCR reaction system, and the reaction system is the same as that described in Example 1.

[0043] Use primers PjF1 / PjR1 to determine the minimum detection amount, and use different dilutions of the mole beetle genome DNA as a template for PCR amplification, and the detection limit of DNA concentration is 0.00585938ng / μL ( figure 2 ).

[0044] 2) Preparation of the mole beetle plasmid

[0045] According to the implementation example 1, the genomic DNA of the mole beetle was amplified, and afte...

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Abstract

The invention relates to Propylaea Japonica specific COI primers and a detection kit containing the primers. According to a unique mtDNA sequence of Propylaea Japonica, a pair of Propylaea Japonica specific COI primers, including PjF1 and PjR1 (SEQ ID No. 1 and 2), is designed, the pair of primers just has a specific amplification ability to Propylaea Japonica, the size of an amplification product is 256bp, the concentration detection limit of plasmids is 1.205943134 E+04, and the concentration detection limit of DNA is 0.00585938ng/mu L. According to the invention, a PCR technology is adopted, so that the detection accuracy is improved, and the detection time is saved, therefore, the Propylaea Japonica specific COI primers and detection kit containing the primers disclosed by the invention are suitable for popularization and application at the local level.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a molybrush-specific COI primer, a kit containing the primer and a detection method thereof. Background technique [0002] Propylea japonica (Thunberg) is a predatory natural enemy insect of the family Coleoptera. It is commonly found in farmland weeds and orchard bushes, and is widely distributed. The tortoise beetle can prey on ladybugs, leafhoppers, planthoppers and other pests, with a large amount of predation; and the tortoise beetle has a high population density in the field, and it is the dominant predatory ladybug in my country's farmland ecosystem Insects play an important role in the natural control of crop pest populations. [0003] With the in-depth research on the predation behavior of ladybugs, it is found that there is a common phenomenon of intragroup predation among ladybugs, and this phenomenon has a complex and huge impact on the population dynamics of differen...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 杨帆王倩陆宴辉徐建祥
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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