New combination of universal amplification primers for plant parasitic nematode ribosome ITS (internal transcribed spacer) and application method thereof

Abstract

The invention provides a new combination of two pairs of universal amplification primers for plant parasitic nematode ribosome ITS (internal transcribed spacer) and an application method thereof for PCR (polymerase chain reaction) amplification. The key innovation point of the method is that after the new primer combination is applied to the PCR amplification of the plant parasitic nematode ITS, the plant nematode type which can be amplified by the new PCR amplification system has broader spectrum, the amplification efficiency and success rate are remarkably improved, and the problems of the ITS amplification in the plant nematode identification before are effectively solved. The PCR amplification product amplified by the method provided by the invention meets the needs of the molecular detection technologies such as nematode DNA (deoxyribonucleic acid) barcode identification and RFLP (restricted fragment length polymorphism) enzyme digestion identification, and can be popularized to the quarantine identification of the port and agriculture/forestry departments in China.

Description

technical field [0001] The present invention relates to the field of quarantine and identification of plant nematodes, and provides two new combinations of universal amplification primers for the ribosome ITS region of plant parasitic nematodes and a method for using them. Specifically, the method uses the new combination of general primers for PCR amplification of the ITS region of the plant parasitic nematode , significantly improved the amplification efficiency and success rate of the established PCR system, and the species of plant parasitic nematodes that can be amplified are also more broad-spectrum, effectively solving many problems in the previous expansion of the ITS region of plant parasitic nematodes, suitable for Application in quarantine and identification laboratories related to ports and agriculture and forestry. Background technique [0002] Accurate identification of plant nematodes is an important basis for the study of nematode biological learning and path...

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is to overcome the defects of low efficiency, low success rate, and low broad-spectrum in the amplification of the ribosome ITS region of the plant parasitic nematode in the prior art, and provide 2 pairs of PCR amplification of the ribosome ITS region of the plant parasitic nematode. The primer combination and the method of use thereof for augmentation, the steps of the primer combination and the method of use are as follows:

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