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Method for rapidly identifying coelomactra antiquate colony by rDNA ITS2 (recombinant Deoxyribose Nucleic Acid Internal Transcribed Spacer) sequence tag

A Xishi tongue and population technology, which is applied in the field of rapid identification of Xishi tongue population by rDNA ITS2 sequence markers, can solve the problems of lack of molecular marker identification methods, difficulties in group identification, adverse effects, etc., and achieve rapid, efficient and accurate identification, and avoid confusion and variation. The effect of fewer sites

Inactive Publication Date: 2014-01-08
INST OF OCEANOLOGY & MARINE FISHERIES JIANGSU
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AI Technical Summary

Problems solved by technology

Due to the similar phenotypes of Shih Tzu tongues in different populations, it is quite difficult to identify populations based on phenotypes
With the introduction of artificially cultivated seedlings from other places, it is easy to cause confusion and adverse effects on the germplasm to a certain extent; how to distinguish seedlings from different regions and the selection of parent shellfish in the later stage has become an important factor for the protection of Xishi tongue germplasm resources and the selection of improved varieties. important issues in education
[0004] There is currently no molecular marker identification method for different populations of Shih Tzu tongue

Method used

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  • Method for rapidly identifying coelomactra antiquate colony by rDNA ITS2 (recombinant Deoxyribose Nucleic Acid Internal Transcribed Spacer) sequence tag
  • Method for rapidly identifying coelomactra antiquate colony by rDNA ITS2 (recombinant Deoxyribose Nucleic Acid Internal Transcribed Spacer) sequence tag
  • Method for rapidly identifying coelomactra antiquate colony by rDNA ITS2 (recombinant Deoxyribose Nucleic Acid Internal Transcribed Spacer) sequence tag

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Embodiment 1

[0023] 1) Sample collection and DNA extraction from different sources:

[0024] Naturally grown Xishi tongue individuals were collected from sea areas such as Jimo (JM) and Rizhao (RZ) in Shandong Province, Ganyu (GY) and Qidong (QD) in Jiangsu Province, and Changle (CL) in Fujian Province. The adductor muscle tissue of the sample was taken with a sterilized scalpel and forceps. The total genomic DNA can be extracted by DNA extraction kit or phenol-chloroform extraction method. Finally, the OD260 and OD280 values ​​of the sample DNA were measured by UV spectrophotometer to determine its concentration and purity, and the mother solution was stored at -20°C. The DNA concentration of 100ng / ul was stored at 4°C for use.

[0025] 2) PCR amplification conditions:

[0026] The primer base sequences are:

[0027] ITS2-F(2A): 5′-GGG TCG ATG AAG AAC GCA G-3′

[0028] ITS2-R(2B): 5′-GCT CTT CCC GCT TCA CTC G-3′

[0029] PCR reaction system is 30μL, each reaction system contains: 50...

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Abstract

The invention relates to a method for rapidly identifying a coelomactra antiquate colony by an rDNA ITS2 (recombinant Deoxyribose Nucleic Acid Internal Transcribed Spacer) sequence tag. The method comprises the steps of (1), collecting a coelomactra antiquate individual, and extracting DNA; (2), synthesizing an ITS2 sequence tag primer of the rDNA; (3), carrying out PCR (Polymerase Chain Reaction) amplification of the ITS2 sequence on a coelomactra antiquate colony sample; (4), purifying PCR products; and (5), carrying out sequencing and sequence alignment on the purified PCR products to obtain a specific locus, and judging the type of the sample. The method is stable in result, high in repeatability and simple in operation, and is of great importance in genetic resource conservation and scientific utilization of shellfish.

Description

technical field [0001] The invention relates to the field of identification of Xishi tongue germplasm, in particular to a method for rapid identification of Xishi tongue population by rDNA ITS2 sequence marker. Background technique [0002] Coelomactra antiquata (Spengler, 1802) is a member of the mollusca (Lamellibranchia), Heteridonta, Veneroida, Mactridae, Coelomidae ( Coelomactra) shellfish, Chinese common names are: "sea mussel", "red egg", "guifei mussel", etc.; the shell is slightly triangular in shape, the shell is thin and brittle, and the shell surface is pinkish-purple near the top; in the natural sea area, the 1-year-old shell The length is 4 to 6cm and the weight is about 20g; the 2-year-old shell can be up to 8cm long and weigh about 110g; the 3-year-old shell can be more than 10cm long and weigh more than 150g, and the larger one can reach more than 250g. Xishi tongue is not only large in size, tender in meat and delicious in taste, but also contains a variet...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q2600/156
Inventor 陈淑吟吉红九陆勤勤张美如
Owner INST OF OCEANOLOGY & MARINE FISHERIES JIANGSU
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