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Universal single primer, kit and method for identifying tridacna crocea, tridacna derasa and hippopus hippopus

A technology of Tridacna safranus and a single primer, which is applied in biochemical equipment and methods, microbe measurement/inspection, DNA/RNA fragments, etc., can solve the problems of differentiation and small difference in gene sequence, and achieve low cost and simple method , the result is intuitive

Active Publication Date: 2020-10-13
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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AI Technical Summary

Problems solved by technology

In recent years, molecular biology based on differences in COI and 16S rDNA mitochondrial gene sequences has been used in the identification of giant clam species, but the differences between these gene sequences are relatively small, and it is difficult to distinguish them by simple methods such as electrophoresis

Method used

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  • Universal single primer, kit and method for identifying tridacna crocea, tridacna derasa and hippopus hippopus
  • Universal single primer, kit and method for identifying tridacna crocea, tridacna derasa and hippopus hippopus

Examples

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Embodiment 1

[0022] See figure 1 , Use sterilized tweezers and small scissors to cut a small amount of tissue from the mantle of Saffron crustacean, non-scale crustacean, and clam oyster. Add 400 μl of lysate and 10 μl of proteinase K (10 mg / ml) to the centrifuge tube containing the mantle tissue, mix on a shaker, and digest in a water bath at 55° C. until the tissue is completely dissolved to obtain the first solution. Add an equal volume of saturated phenol (200μl), chloroform / isoamyl alcohol (24:1) (200μl) mixture for extraction three times, centrifuge at 10000rpm for 10-15min each time, and aspirate the supernatant with a pipette. Add 1 mL of pre-cooled absolute ethanol to the centrifuge tube containing the supernatant, and settle overnight. Finally, it was washed with 70% alcohol, dried at room temperature, and dissolved in 200 μL of ultrapure water. UV spectrophotometer was used to detect DNA concentration and purity, and 1% agarose gel electrophoresis was used to detect DNA integri...

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Abstract

The invention discloses a universal single primer, kit and method for identifying tridacna crocea, tridacna derasa and hippopus hippopus. The method for detecting and identifying the tridacna crocea,the tridacna derasa and the hippopus hippopus by using universal single primer multiplex PCR of competitive annealing of multiple downstream primers is established, and actual sample tests prove thatthe method has the advantages of being simple, intuitive in result, accurate, effective, free from influences of environment and development period, low in cost and the like.

Description

Technical field: [0001] The invention belongs to the technical field of molecular identification in aquatic products, and specifically relates to a universal single primer, a kit and an identification method for the identification of saffron clams, non-scale clams and clams. Background technique: [0002] The universal single-primer multiplex PCR technology is to add a universal primer and multiple species-specific primers to a PCR reaction system. Only the sequence paired with the specific primer can be amplified, separated by agarose gel electrophoresis, and then Compare with standard fragments to realize species identification. The universal single-primer multiplex PCR technology has the advantages of high efficiency, rapidity, and reliable results, and is widely used in species identification. [0003] The clams belong to the phylum Mollusca, Phytobranchia, Clamidae, and Clams. It is a large tropical marine coral reef benthic shellfish and the largest bivalve in the ocean. Th...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/6888C12Q1/686C12Q2600/16C12Q2537/143C12Q2565/125
Inventor 马海涛高红梅喻子牛张跃环
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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