Method for preparing fish ovary germ cell chromosome

Abstract

The invention discloses a method for preparing a fish ovary germ cell chromosome. The method comprises the following steps of: preparing a living mature ovary and washing the living mature ovary with normal saline; putting the ovary in normal saline containing 1mug / ml estrogen, cutting the ovary into small sections, and putting the ovary sections on a shaker for culturing for 1-3 hours at room temperature by keeping out of the sun; coating a layer of 1 percent agar at the bottom of a culture dish and filling a 4 percent glacial acetic acid solution; putting several cultured ovary sections into the 4 percent glacial acetic acid solution; after the ovary sections move to an animal pole, peeling off the ovokaryons; peeling off yelks on the peripheries of the ovokaryons; putting the ovokaryons in pre-cooled Kano stationary liquid for freezing overnight, wherein the Kano stationary liquid is prepared by mixing methanol and glacial acetic acid together, and the volume ratio of the methanol to the glacial acetic acid is 3:1; putting 3-5 ovokaryons frozen overnight on a glass slide for 10-20 seconds; then putting the glass slide carrying the ovokaryons into a dye vat containing a coloringagent for dying for 30-45 minutes by keeping out of the sun; and after the glass slide is soaked for 30 minutes in pure water by keeping out of the sun, covering cover glass on the glass slide.

Description

Technical field: [0001] The invention belongs to the technical field of cytogenetics, in particular to a preparation of fish ovary germ cell chromosomes which is simple and fast in operation method and can clearly observe the number of chromosomes in the meiosis stage of fish egg nuclei and synapsis under a fluorescent microscope method. Background technique: [0002] Natural polyploidy occurs in fish. Natural polyploid fish has the characteristics of fast growth, disease resistance, and high nutritional value, and has important germplasm value in genetic breeding; natural tetraploid fish and diploid fish can be crossed to obtain 100% triploid, The sterility of triploid fish is also of great significance for controlling the overproduction of farmed fish and protecting natural germplasm resources; at the same time, chromosome manipulation can be used to induce higher ploidy fish (pentaploid, etc.), using The fertilization of the genetically inactivated sperm and the natural...

AI Technical Summary

Problems solved by technology

There are also a small number of early-developed ovaries, but because most oogonia are mainly mitotic, it is difficult to observe the meiotic phase; and mature ovaries are disturbed by yolk and other accessory structures, which give the oocyte chromosomes a band come very difficult

So far, there has been no report on extracting egg nuclei from egg cells to prepare chromosomal phases

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