PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) method for identifying seven sea cucumber species

Abstract

The invention discloses a PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) method for identifying seven sea cucumber species. The PCR-RFLP method includes the steps of firstly extracting DNA (deoxyribonucleic acid) modules of sea cucumbers; subjecting specificity fragment of a CO I (coenzyme I) gene to PCR amplification to obtain a fragment about 690bp; secondarily subjecting amplified product to restriction enzyme digestion; and finally subjecting the enzyme-digested product to electrophoresis detection. Different sea cucumbers obtain stripes in different numbers and sizes, and selected specificity incision enzymes respectively include: Californian paracanthurus cucumber: SSP I, North Atlantic Ocean cucumaria: EcoR V, apostichopus japonicas: SSP I, thelenota ananas: EcoR V, acaudina molpadioidea: Sca I, isostichopusbadionotus: Pvu II, and actinopyga mauritiana: EcoR V and SSP I. The PCR-RFLP method overcomes the defects that a conventional morphological identification method is prone to affecting in accuracy, is heavy in workload and long in consumption time and the like. The PCR-RFLP method can be completed by only one-time PCR reaction, one-time enzyme digestion reaction and one-time polypropylene gel electrophoresis, and is simple to operate, short in parting time and good in specificity.

Description

technical field

[0001] The invention relates to a method for identifying sea cucumber species in molecular biology, in particular to a PCR-RFLP method for identifying seven sea cucumber species. Background technique

[0002] Sea cucumber is a traditional marine food and medicine resource in my country. Due to the large differences in the nutritional value and economic value of different types of sea cucumbers, some unscrupulous traders often deceive consumers by using false ones and shoddy ones. Sea cucumbers that have lost their original form are more prone to counterfeiting.

[0003] The traditional sea cucumber species identification method is morphological identification, which is mainly based on the external morphology and anatomical characteristics of sea cucumbers: external characteristics include the shape of tentacles, the presence or absence of warty feet and tube feet, their shape and distribution characteristics, etc.; anatomical characteristics mainly refer to ...

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