Method for identifying DNA bar code molecule of earthworm

Abstract

The invention discloses a method for identifying a DNA bar code molecule of an earthworm and a COI sequence of the DNA bar code molecule of the earthworm. The method comprises the steps of amplifying the gene of a sample COI through polymerase chain reaction (PCR), verifying the PCR product by agarose gel, then sending to a biological sequencing company for sequencing, according to the sequencing result, sequence splicing through manual correcting, comparing with a public sequence, then confirming the tissue to be tested as the source of pheretima aspergillum or pheretima vulgaris if the homology of the spliced sequence and the gene sequence SEQ ID NO.1-NO.4 is over 99%. By adopting the reliable DNA molecular identification technology, the method can rapidly and accurately identify the earthworm varieties specified by the 'pharmacopoeia of China' and non-medicinal material earthworm varieties without conducting morphological identification on the earthworm varieties, can further distinguish the pheretima aspergillum varieties from pheretima vulgaris varieties, improves the accuracy and reliability, and guarantees the safety of the earthworm as the medicinal material.

Description

technical field [0001] The invention relates to the field of species identification, in particular to a DNA barcode molecular identification method of earthworm. Background technique [0002] Earthworm, that is, earthworm, is cold in nature and slightly salty in taste. It is a traditional Chinese medicinal material. However, only the annelid phylum Megaterridae is recorded under the item "Earth Dragon" in the Pharmacopoeia of the People's Republic of China 2010 Edition (Part One). Dried body of Pheretima aspergillum (E. Perrier), Pheretima vulgaris Chen, Pheretima guillelmi (Michaelsen) or Pheretima pectinifera Michaelsen. The former is commonly called "Guangdilong", and the latter three are commonly called "Shanghaidilong". Earthworm is listed as inferior in "Shen Nong's Materia Medica", which has the effects of clearing away heat and calming convulsions, dredging collaterals, relieving asthma, and diuresis. It is often fried and used for high fever, coma, convulsions, ar...

AI Technical Summary

Problems solved by technology

However, since most of the animal medicinal materials have similar external shapes and non-specific tissue characteristics, and after rough processing in the place of origin and processing of decoction pieces, it is difficult to see the shape and color of the species itself, and the microscopic characteristics are also damaged to varying degrees, resulting in the dry product of Dilong. It is difficult to identify

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