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A method for identifying blue crabs cultured in Sanmen area
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A mud crab and fragment technology, which is applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as instability and difficulty in distinguishing
Active Publication Date: 2021-04-09
NINGBO UNIV
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However, it is not easy to distinguish from the appearance, and it is also very unstable, especially through chemical treatment to make the appearance the same
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Embodiment 1 3
[0024] Example 1 Screening of the characteristic DNA sequence of intestinal microorganisms of the mud crab in Sanmen area
[0025] 1) Collect samples of mud crabs from different regions
[0026] Sanmen blue crab is a pseudo-cave blue crab in biological classification, and its natural distribution and cultivation are mainly in the southeast coast of my country, including Zhejiang, Fujian, Guangdong, Guangxi, and Hainan. Collection of mud crabs from different regions. At the same time, according to the concentrated (massive) listing time of Sanmen blue crabs in Zhejiang, they are divided into around the Dragon Boat Festival, around the Mid-Autumn Festival, and from the end of November to the end of the year. Therefore, three batches of sample collection were carried out for the time of centralized listing.
[0027] 2) Intestinal microbial analysis
[0028] The intestines and contents of 9 female and 9 male crabs in a single batch of sampling points were collected, and the sam...
Embodiment 2
[0049] Embodiment 2 is used for identifying the kit of mud crab
[0050] 1) Composition of the kit
[0051] The composition of the kit includes SMO, SM_sF, SM_sR, SM_uF, SM_uFR, SM1. SM0 is DNA in vitro amplification premix liquid, the main components are PCR reaction butter, dNTP and Taq enzyme, SM1 is ddH2O. SM_sF and SM_sR are respectively forward and reverse primers (SEQ ID NO:5 and SEQ ID NO:6) of specific primers; SM_uR and SM_uF are respectively forward and reverse primers of universal primers (SEQ ID NO:2 and SEQ ID NO:2 and SEQ ID NO:6). NO: 3).
[0052] The method of using the kit is as follows:
[0053] 1. DNA extraction from mud crab intestines and excreta
[0054] Please refer to the DNA extraction kit (DNeasy Kit). The specific operation is as follows:
[0055] 1. Add 0.25g soil sample to a PowerBead Tubes;
[0056] 2. Gently vortex to mix
[0057]3. Test Solution C1. If precipitation occurs, take a water bath at 60°C until completely dissolved
[005...
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Abstract
The present invention provides an identification method for blue crabs, which is determined by detecting whether there are nucleotide fragments of microorganisms belonging to the genus Psychrilyobacter in the intestinal microorganisms of blue crabs; the nucleotide fragments are 16S rDNA fragments, and the nuclear The nucleotide sequence is SEQ ID NO:4. The present invention collects blue crabs from the main producing areas of blue crab breeding in the southeast coastal provinces of China, and finally screens out the specific microbial populations of blue crabs, and comprehensively designs the characteristic DNA sequences of these populations to obtain the "molecular" for identifying blue crabs. "Fingerprint" and developed into a kit, thus solving the difficult problem of identification of Sanmen blue crab.
Description
technical field [0001] The invention belongs to the technical field of crab identification, and in particular relates to a method for identifying blue crabs cultured in Sanmen area. Background technique [0002] The types of marine crabs are very rich in resources. There are more than 2,000 species of crabs in the Indo-Pacific region alone, and the crabs in my country account for about one-third of them. At present, there are only three species of crabs that are artificially bred in a large scale in my country: Chinese mitten crab (Eriocheir sinensis), portunus trituberculatus (Portunus trituberculatus) and blue crab (Scyllaparamamosain). Among them, the Chinese mitten crab is a freshwater crab, and the latter two are seawater crabs. The former two are distributed in the coastal areas of my country, while the blue crab is distributed in the tropical and subtropical southeast coast of my country. The most famous one is the Sanmen blue crab in Zhejiang, which enjoys the reput...
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