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An identification method and a technology for the spinnaker mussel, applied in the field of molecular biology, can solve the problems of low accuracy and large limitations, and achieve the effects of quick and simple judgment, simple and intuitive results, and improved detection efficiency.
Pending Publication Date: 2021-03-12
SHANGHAI OCEAN UNIV
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However, this method is based on the judgment of human experience and is suitable for experienced personnel, and this method can only be targeted at mature and in the breeding period of the triangle sail, the limitations are relatively large, and the accuracy of the identification is not good. high
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Embodiment 1
[0037] The identification method of the sex of the juvenile triangular sail clam of the present embodiment comprises the steps:
[0038] 1. Sample Collection
[0039] Randomly select 20 4-month-old and 5-month-old Trigonopsis mussels, use different sets of tweezers, scissors and other sampling tools to avoid cross-contamination, take a small amount of gonads, put them into a 1.5mL sterilized centrifuge tube, and quickly freeze them in liquid nitrogen , used directly for DNA extraction after sampling.
[0040] 2. DNA extraction and concentration determination
[0041] (1) Add 500 μL of phenol, turn it upside down for 8 minutes, centrifuge at 11000 r / min at 6°C for 10 minutes.
[0042] (2) Aspirate the supernatant, add phenol:chloroform:isoamyl alcohol at a volume ratio of 25:24:1, invert upside down for 8 minutes, centrifuge at 11000 r / min for 10 minutes at 6°C.
[0043] (3) Aspirate the supernatant, add chloroform:isoamyl alcohol with a volume ratio of 24:1, invert upside d...
Embodiment 2
[0053] The method for identifying the sex of the mature Triangle sail clam of the present embodiment comprises the following steps:
[0054] 1. Sample collection
[0055] Randomly select 44 second-year-old triangular sailfish, use different sets of tweezers and scissors and other sampling tools to avoid cross-contamination, take a small amount of gonads and put them into a 1.5mL sterilized centrifuge tube, quickly freeze them in liquid nitrogen, and use them directly after sampling. for DNA extraction.
[0056] 2. DNA extraction and concentration determination
[0057] (1) Add 500 μL of phenol, turn it upside down for 8 minutes, centrifuge at 11000 r / min at 6°C for 10 minutes.
[0058] (2) Aspirate the supernatant, add phenol:chloroform:isoamyl alcohol at a volume ratio of 25:24:1, invert upside down for 8 minutes, centrifuge at 11000 r / min for 10 minutes at 6°C.
[0059] (3) Aspirate the supernatant, add chloroform:isoamyl alcohol with a volume ratio of 24:1, invert upside...
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Abstract
The invention provides a hyriopsis cumingii gender identification method, and the method comprises: collecting a small amount of gonad tissue samples, and extracting DNA; designing a primer accordingto an excessive specific sequence of the M-type COII gene for PCR amplification; finally, observing strip results through agarose gel electrophoresis to judge male and female hyriopsis cumingii, and judging the hyriopsis cumingii with strips as male hyriopsis cumingii and the hyriopsis cumingii without strips as female hyriopsis cumingii. According to the method, the gender of hyriopsis cumingii is accurately identified mainly according to the genome amphiphilic and monophilic heredity phenomenon of hyriopsis cumingii and the characteristic that COII genes in an M-type mitochondrial genome arespecifically expressed in male gonads, and the result is simple and intuitive, so that the gender of hyriopsis cumingii is accurately identified; therefore, the method can be used for effectively distinguishing genders of early-stage fry of hyriopsis cumingii, and is less limited by samples and high in accuracy; besides, the DNA sample is used for judging more quickly and simply through gel electrophoresis, a subsequent sequencing result is not needed, and the detection time is greatly shortened, so that the detection efficiency is improved.
Description
technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a method for identifying the sex of the triangular sailfish. Background technique [0002] Triangle clams, commonly known as river mussels, pearl mussels, and triangle mussels, are a kind of freshwater bivalve mollusks, belonging to the clambranchial class, the euplobranchial order, the pearl clam family, and the genus Sail. It is the main freshwater pearl breeding mussel in China, and more than 80% of domestic freshwater pearls are bred by it. In sexually mature Trigonopsis, there are significant differences in pearl-producing performance between male and female, and males have better pearl-producing ability. Selecting the male Triangle sail clam for refined cultivation is of great significance to the improvement of pearl production capacity. However, in actual production, sex identification of male and female mussels is still a difficult problem. From a...
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