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Detection method for DNA damage of embryos of sepiella maindroni

A Mansoni-free cuttlefish and DNA damage technology, which is applied in the field of DNA damage detection, can solve the problems of tumor formation, DNA damage, cell apoptosis, etc. in the body, and achieve the effect of meeting the detection amount, small sample size, and good repeatability

Inactive Publication Date: 2019-01-04
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, during the process of self-replication, transcription and exposure to various external environments, biological organisms are extremely susceptible to DNA damage caused by internal or external factors. Under normal circumstances, most of the damage can be recognized and repaired by cells. However, when the DNA damage is serious or the DNA damage repair ability of the body is lacking, and the body cannot repair normally, the DNA with this damage will enter a semi-conservative replication process, and base mismatches are prone to occur, thereby causing cell apoptosis and even cell death. Promotes the formation of tumors in the body, as well as resistance to various types of cancer cells

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] A method for detecting DNA damage of squid squid embryos, comprising the collection of fertilized eggs and embryos at different development stages and the detection of DNA damage, specifically comprising the following steps:

[0019] 1) Collection of fertilized eggs and embryos: select sexually mature cuttlefish with strong swimming and healthy individuals, and the ratio of male to female is 1:1.5. They are mixed and reared at 25°C. After overnight in the dark, they are stimulated by light the next morning. Male and female fish mate naturally to lay eggs, collect fertilized eggs and embryos of different development stages, wash them quickly with PBS, and select embryos of each development stage under a dissecting microscope;

[0020] 2) Detection of DNA damage: Rinse the embryo twice with PBS, peel off the egg membrane, and digest it with a digestive solution containing proteinase K and salicylic acid at room temperature. After electrophoresis, the slides were taken out...

Embodiment 2

[0028] A method for detecting DNA damage of squid squid embryos, comprising the collection of fertilized eggs and embryos at different development stages and the detection of DNA damage, specifically comprising the following steps:

[0029] 1) Collection of fertilized eggs: Select sexually mature squid with strong swimming and healthy individuals, and the ratio of male to female is 1:2.0. They are mixed and reared at 26°C. After overnight in the dark, the light stimulates the male and female fish the next morning. Natural mating to lay eggs, collect fertilized eggs, wash them quickly with reconstituted water, and select embryos with normal development and blastocyst stage after fertilization under a dissecting microscope;

[0030] 2) Detection of DNA damage: Rinse the embryos twice with PBS, peel off the egg membrane, and digest with a digestion solution containing 10 µg / ml proteinase K and 1.2 µg / ml salicylic acid at room temperature, and digest the embryos that have developed...

Embodiment 3

[0035] A method for detecting DNA damage of squid squid embryos, comprising the collection of fertilized eggs and embryos at different development stages and the detection of DNA damage, specifically comprising the following steps:

[0036] 1) Collection of fertilized eggs: Select sexually mature squid with strong swimming and healthy individuals. The ratio of male to female is 1:2.5, and they are mixed at 28°C. After overnight in the dark, light stimulates the male and female fish the next morning. Natural mating to lay eggs, collect fertilized eggs, wash them quickly with reconstituted water, and select embryos with normal development and blastocyst stage after fertilization under a dissecting microscope;

[0037] 2) Detection of DNA damage: Rinse the embryos twice with PBS, peel off the egg membrane, and digest with a digestion solution containing 12 µg / ml proteinase K and 1.5 µg / ml salicylic acid at room temperature, and digest the embryos that have developed for ≤24 hours ...

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Abstract

The invention discloses a detection method for DNA damage of embryos of sepiella maindroni. The method comprises the following steps of selecting sepiella maindroni which is high in mobility, healthyand sexually mature, making sepiella maindroni naturally mate and spawn, collecting fertilized eggs, and selecting the embryos in different development stages after fertilization; rinsing the embryoswith PBS twice, peeling off egg membranes, and adopting a digestive solution containing proteinase K and salicylic acid for digestion at room temperature; after complete digestion, utilizing agarose to make a glue plate, then putting the glue plate in a pyrolysis solution for pyrolysis, conducting electrophoresis and dyeing, observing an image under a fluorescence microscope, and conducting measurement. The detection method has the unique advantages that the method is simple, quick, low in consumption, high in sensitivity, accuracy and reproducibility, low in cost and the like, few samples arerequired, the coloring effect is good, and the complete and clear image can be obtained and is easy to analyze.

Description

technical field [0001] The invention relates to the technical field of DNA damage detection, in particular to a method for detecting DNA damage of squid squid embryos. technical background [0002] Cephalopods belong to molluscs, which are an important part of the marine ecosystem and play an indispensable role in the balance of marine ecology. Squid belongs to the phylum Mollusca and the class Cephalopoda. It is widely distributed in various sea areas and plays an important role in the evolution of biological systems. Its meat is delicious and delicious, with good nutritional value and high protein content. In addition to being edible, it also has high medicinal value. Ink sacs can promote blood coagulation and have antibacterial functions. Sea octopuses can be used for medicine. Otoliths can be used to analyze the age of species and the temporal and spatial changes of the marine environment. In addition, squid plays a very important role in the food chain of the marine e...

Claims

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Application Information

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IPC IPC(8): G01N27/447G01N21/64G01N1/30
CPCG01N27/447G01N1/30G01N21/6458
Inventor 柳意樊刘炳舰张建设景斐夏立萍
Owner ZHEJIANG OCEAN UNIV
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