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Acquisition method of substance of cell nucleus

A technology for obtaining methods and nuclear substances, which can be applied in the fields of botanical equipment and methods, biochemical equipment and methods, and plant genetic improvement, etc., can solve problems such as prokaryotic rupture, and achieve the effect of easy operation and easy popularization and use.

Active Publication Date: 2013-05-22
SHANDONG UNIV +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, this method has some disadvantages as follows: 1. Electric fusion or other fusion methods must be used to transfer the new nucleoplasmic body to the new recipient cytoplasmic body; 2. Prokaryotic cells are easily squeezed by the cytoplasm during the absorption This is unacceptable in future clinical applications; 3. It is inevitable to bring in part of the cytoplasm

Method used

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  • Acquisition method of substance of cell nucleus
  • Acquisition method of substance of cell nucleus
  • Acquisition method of substance of cell nucleus

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Embodiment

[0026] (1) Use a laser to make a hole with a diameter of 30 μm in the larger zona pellucida part of the perivitelline space of the GV stage oocyte, and the laser frequency is 1.8 ms;

[0027] (2) Fix the oocyte with an egg-holding needle with an outer diameter of 120 μm and an inner diameter of 25 μm, so that the opening end of the zona pellucida is located opposite to the egg-holding needle, wherein the opening of the egg-holding needle is flat;

[0028] (3) Insert 1 μl of G1.5 culture medium containing 5% HSA into the cytoplasm from the opening of the zona pellucida with an injection needle with an inner diameter of 40 μm, and avoid direct contact with GV vesicles;

[0029] (4) Rapidly increase the positive pressure in the injection needle to make the liquid in the needle flow outward quickly, so that the liquid in the injection needle can quickly break the connection between the GV vesicle and the cytoplasm, and use the increase in the positive pressure in the entire cell to...

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Abstract

The invention, belonging to the technical field of biology, relates to an acquisition method of the substance of a cell nucleus for a clone technology, comprising the following steps: (1) perforating on the zona pellucid portion with large perivitelline space by laser; (2) fixing the cell through an ovum holding pin to let the open end of the zona pellucid be opposite to the ovum holding pin; (3)pushing an injection needle with liquid into cytoplasm from the opening of the zona pellucid and avoiding the direct contact with the substance of the cell nucleus; (4) rapidly increasing positive pressure in the injection needle to allow the liquid in the injection needle rapidly flow outwards to push and cut the connection between the substance of the cell nucleus and the cytoplasm to make no cytoplasm around the substance of the cell nucleus, and using the increasement of the positive pressure in the whole cell to allow the substance of the cell nucleus completely to be removed from the cell from the opening of the zona pellucid; and (5) using the injection needle to collect the separated substance of the cell nucleus. The method has no need of special devices, can realize the separation of the substance of the cell nucleus simply and conveniently, and can avoid the facture of the substance of the cell nucleus, and the obtained substance of the cell nucleus is convenient for cell fusion experiments.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to a method for transferring cell genetic material, more specifically to a method for obtaining cell nucleus material. Background technique [0002] As the genetic material, the nuclear material determines the genetic characteristics of organisms, and the nucleus is the main gathering place of genetic material. Pronuclear stage embryos have a clear pronucleus surrounded by a complete nuclear membrane, which can be accurately observed with an inverted microscope without any auxiliary equipment and reagents, which provides the necessary prerequisites for the precise acquisition and transfer of genetic material. The currently used prokaryotic acquisition method is mainly the biopsy needle suction cytoplasm method, that is, the biopsy needle is used to absorb the cytoplasm near the prokaryotic nucleus, and finally the prokaryotic nucleus together with part of the cell membrane and cytoplasm fo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/873
Inventor 陈子江吴克良马金龙
Owner SHANDONG UNIV
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