Method for identifying helix pomatia by PCR (polymerase chain reaction) detection

Abstract

The invention discloses a method for identifying helix pomatia by PCR (polymerase chain reaction) detection, which uses specific primers to identify molecular characteristics of helix pomatia. The forward primer for the PCR reaction is HP-(P1): 5'-TTCGTTGAGAGTTAGG-3', and the reverse primer is HP-(P2): 5'-ATAGTAATAGCACCAGC-3'. The total volume of the PCR reaction system is 25 mu L, wherein the 2*TaqPCRMasterMix is 12.5 mu L, the forward and reverse primers are respectively 0.5 mu L, the DNA (deoxyribonucleic acid) template is 2 mu L, and the rest is sterilized ddH2O. The PCR reaction process comprises the following steps: pre-denaturating at 5 DEG C for 5 minutes; treating at 95 DEG C for 30 seconds, 45 DEG C for 30 seconds and 72 DEG C for 1 minute, and repeating 30 times; and extending at 72 DEG C for 10 minutes, and finishing the reaction. The method disclosed by the invention can quickly and accurately detect and identify helix pomatia. The invention provides a new technical means for detection and identification of helix pomatia in the Catalogue of Quarantine Pest for Import Plants to the People's Republic of China, and has important meanings for preventing helix pomatia from spreading and diffusion.

Description

technical field [0001] The invention relates to the field of molecular biology detection and identification, in particular to a method for PCR detection and identification of large snails. Background technique [0002] cover big snail Helix pomatia (Linnaeus, 1758) is a member of the "List of Imported Plant Quarantine Pests of the People's Republic of China" issued by my country in 2007. Helix aspersa (Müller), bright big snail Helix lucorum (Linnaeus, 1758), etc. are very similar, often difficult to distinguish, and only senior terrestrial mollusc taxonomy experts can identify, and the identification of eggs and juvenile snails is even more difficult. Due to historical reasons, the vast majority of plant quarantine officers at border ports are majors in plant protection and know little about the classification of terrestrial molluscs. common technical challenges. In the past two decades, molecular systematics has developed rapidly, that is, the comparison of gene seq...

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Problems solved by technology

[0002]The large cover snail Helix pomatia (Linnaeus,1758) is the "People's Republic of China Entry Plant Quarantine Pests" promulgated in 2007 The members of the list, whose shells are very similar to those of the same genus Helix aspersa (Müller), Helix lucorum (Linnaeus, 1758), etc., are often difficult to identify. Differentiation, and only senior terrestrial mollusc taxonomy experts can identify, the identification of eggs and young snails is more difficult

Due to historical reasons, the vast majority of plant quarantine officers at border ports are majors in plant protection and know little about the classification of terrestrial molluscs. common technical challenges

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