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DNA barcoding standard gene sequence of Rizhao Blepharipoda liberata and species identification method based thereon

A standard gene and gene sequence technology, applied in the field of species identification, can solve the problem of no sunshine sea cicada, and shorten the identification time.

Inactive Publication Date: 2014-07-23
刘丰铭
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are some COI gene sequences of Cicadas in Genbank, but there is no related gene sequence of Rizhao sea cicada

Method used

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  • DNA barcoding standard gene sequence of Rizhao Blepharipoda liberata and species identification method based thereon
  • DNA barcoding standard gene sequence of Rizhao Blepharipoda liberata and species identification method based thereon

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Experimental program
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Embodiment 1

[0022] 1. Collection, preservation and processing of samples to be tested

[0023] The samples to be tested were collected at the seashore of Lunan Seashore Forest Park (Rizhao Dashawa), and a part of the muscle tissue was taken, and then the samples were stored in 95% alcohol or in a freezer at -20°C.

[0024] 2. DNA template preparation

[0025] The DNA in the samples was extracted by the phenol-chloroform method and stored at -20°C for future use.

[0026] 3. Primer synthesis

[0027] Forward primer: 5'-GGTCAACAAATCATAAAGATATTGG-3'

[0028] Reverse primer: 5'-TAAACTTCAGGGTGACCAAAAAAATCA-3'

[0029] 4. PCR amplification

[0030] The PCR reaction system of this embodiment is shown in Table 1.

[0031] Table 1 shows the target gene PCR reaction system of the samples to be tested (50uL system)

[0032] Sterile distilled water 37.75 μl 10×Ex Taq Buffer 5 μl dNTP Mixture (2.5 mM each) 4 μl LCO1490 (10 μM) 2.5 μl HCO2198 (10 μM) ...

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Abstract

Relating to the technical field of species identification, the invention specifically relates to a DNA barcoding standard gene sequence of Rizhao Blepharipoda liberata and species identification method based on the gene sequence. The DNA barcoding standard gene is COI gene, which has a gene sequence shown as SEQIDNO.1. The identification method includes: conducting extraction separation of DNA on a to-be-tested tissue, and amplifying a target gene by polymerase chain reaction. A pair of primers used in the amplification reaction are shown as the following: a forward primer 5'-GGTCAACAAATCATAAAGATATTGG-3', and a reverse primer 5'-TAAACTTCAGGGTGACCAAAAAATCA-3'. The gene sequence obtained in the invention is in favor of realizing molecular identification of the Rizhao Blepharipoda liberata, and can effectively shorten the identification time of the Rizhao Blepharipoda liberata.

Description

technical field [0001] The invention relates to the technical field of species identification, in particular to a DNA barcode standard gene sequence of Rizhao sea cicada and a species identification method based on the gene sequence. Background technique [0002] The sea cicada in Rizhao is a kind of creature commonly known as "Hai Zhile" in Rizhao. It looks like a young cicada on land. Its scientific name is Jiefang Meiped Crab, also known as sea cicada crab. marine crustaceans. In the coast of mainland China, it is only produced in the coastal area of ​​Rizhao, Shandong, and is an authentic Rizhao specialty. Sea cicada crabs are crispy and delicious after being fried. Their meat quality is comparable to that of lobsters. In recent years, because the liberated eyebrow crab has been served on the dining table as a seafood specialty, the consumer demand for the liberated eyebrow crab has greatly increased, which has led to an increase in its excavation; in addition, the dis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12Q1/68
Inventor 刘丰铭
Owner 刘丰铭
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