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Specific primers for amplifying Verticillium lecanii

A specific technology for Verticillium cereus, applied in the field of specific primers for amplifying Verticillium cereus

Inactive Publication Date: 2012-07-04
谢明
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention aims to solve the current problem of lacking primers to amplify the target sequence of Verticillium lecanii from sample DNA, and provides a specific primer for amplifying Verticillium lecanii

Method used

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  • Specific primers for amplifying Verticillium lecanii
  • Specific primers for amplifying Verticillium lecanii

Examples

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Embodiment 1

[0010] Verticillium lecanii, Verticillium dahliae, Beauveria bassiana, Metarhizium anisopliae, Paecilomyces fumosoroseus , Fusarium oxysporum, Botrytis cinrea and Alternaria alternata were used as templates to verify the specificity of primers VF / VR.

[0011] The PCR amplification system is: 25 μL consists of 0.5 μL DNA template (about 10 ng), 0.5 μL forward primer VF (10 μM), 0.5 μL reverse primer VR (10 μM), 0.5 μL dNTPs (10 mmol / L), 2.5 μL 10×PCR buffer (with MgCl2), 0.2 μl TaqDNA polymerase (5U / μL), and sterilized double distilled water to make up 25 μL. The PCR amplification reaction conditions were: pre-denaturation at 94°C for 3 min, denaturation at 94°C for 30 s, annealing at 55°C for 30 s, extension at 72°C for 45 s, a total of 35 cycles, extension at 72°C for 5 min, and storage at 4°C.

[0012] The sequence amplified in this embodiment is detected by agarose gel electrophoresis, and the detection results are as follows: figure 1 as shown, figure 1 Lane M in the mi...

Embodiment 2

[0014] The DNA of 24 soil samples (collected from Langfang City, Hebei Province and Wuhan City, Hubei Province) was amplified by PCR using primers VF / VR. The PCR amplification system is: 25 μL consists of 0.5 μL DNA template (about 10 ng), 0.5 μL forward primer VF (10 μM), 0.5 μL reverse primer VR (10 μM), 0.5 μL dNTPs (10 mmol / L), 2.5 μL 10× PCR buffer (with MgCl 2 ), 0.2 μl TaqDNA polymerase (5U / μL), and sterilized double distilled water to make up 25 μL. The PCR amplification reaction conditions were: pre-denaturation at 94°C for 3min, denaturation at 94°C for 30s, annealing at 55°C for 30s, extension at 72°C for 45s, a total of 35 cycles, extension at 72°C for 5min, and storage at 4°C.

[0015] The sequence amplified in this embodiment is detected by agarose gel electrophoresis, and the detection results are as follows: figure 2 as shown, figure 2 Middle lane M is the standard BM2000, lanes 1-12 are the amplification results of soil sample DNA in Langfang City, Hebei ...

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Abstract

The invention relates to a pair of specific primers for amplifying Verticillium lecanii. The invention solves the problem of lack of primers for amplifying the Verticillium lecanii target sequence from a sample DNA. The specific primers for amplifying Verticillium lecanii are (5'-CGGCGTCCGGACGCGGACCCAG-3' and VR(5'-CCCCAACGCCGACTTCCCCGAG-3'). The primers provided by the invention can accurately amplify the target segment of Verticillium lecanii.

Description

technical field [0001] The invention relates to a specific primer for amplifying Verticillium lecanii, which can be used for molecular detection of Verticillium lecanii in samples. Background technique [0002] Verticillium lecanii is an entomopathogenic fungus with a wide geographical distribution and host range. It was first discovered by Nivter in Ceylon in 1861. In my country, Sawada Kaneyoshi of Japan collected samples from Taiwan in 1959. Verticillium lecanii was isolated. Verticillium lecanicillium has always belonged to the genus Verticillium, but according to the strain morphological differences and molecular genetic analysis, it is now changed to the genus Lecanicillium, which contains a very rich variety and can infect pathogenic fungi, pests and pathogenic nematodes on a variety of crops. [0003] Verticillium lecanii has been proved to be a kind of microbial insecticide with great potential. Countries such as Europe, America and the former Soviet Union have succ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 谢明张艳军邱卫亮
Owner 谢明
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