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Embryo taking buffer liquid and application thereof

A buffer and solution technology, applied in the field of biochemistry, to achieve the effect of improving harvesting efficiency

Active Publication Date: 2016-11-16
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there is currently no efficient method for harvesting neurula stage pig embryos

Method used

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  • Embryo taking buffer liquid and application thereof

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Embodiment Construction

[0019] Below in conjunction with specific embodiment, the present invention is further explained:

[0020] 1. Prepare the reaction solution:

[0021] The formula of PBS buffer solution is: 0.1mol / L potassium dihydrogen phosphate solution and 0.1mol / L disodium hydrogen phosphate solution are mixed in a volume ratio of 3:7, and the pH is adjusted to 7.2 to complete 1×PBS buffer solution preparation.

[0022] The formula of embryo taking buffer is: add 0.1% BSA dry powder (does not contain Ca) to 1×PBS buffer 2+ , Mg 2+ and phenol red) to complete the preparation of embryo retrieval buffer.

[0023] 2. Efficient methods for harvesting pig embryos:

[0024] (1) Put the embryo extraction buffer in a 37°C water bath to keep the temperature for later use.

[0025] (2) Add 1×PBS buffer solution to a coning culture dish with a diameter of 3.5cm, and place it on crushed ice for later use.

[0026] (3) Under the constant temperature condition of 37°C, move the frozen neurula stage ...

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Abstract

The invention belongs to the field of biochemistry, and relates to an embryo taking buffer liquid and an application thereof; the embryo taking buffer liquid is prepared by the following steps: carrying out water bath of the embryo taking buffer liquid for standby application, and adding a PBS buffer liquid into a culture dish; taking a refrigerated uterine tissue to remove to a constant temperature stage; on the constant temperature stage, placing another culture dish and the uterine tissue, cutting off uterine mesentery, cutting cornua uteri open at a distal fallopian tube, nipping the distal cornua uteri tightly, cutting proximal cornua uteri open, dividing the uterus into two parts, at a cut-open cornua uteri proximal end, injecting the embryo taking buffer liquid, swashing the cornua uteri, starting from a cornua uteri distal end, extruding cornua uteri contents comprising the embryo taking buffer liquid to the cornua uteri proximal end into the culture dish; and removing the culture dish to under a stereoscope, searching embryos, separating the embryos one by one to put in a culture dish having addition of a PBS buffer liquid, rinsing, numbering and recording. The embryos can be efficiently harvested, the vitality of the harvested embryos can be guaranteed, and the harvested embryos can be used for study of cell culture and auxology and thremmatology.

Description

technical field [0001] The invention belongs to the field of biochemistry, and relates to an embryo taking buffer solution and an application thereof. Background technique [0002] Starting from the union of the embryos of the two sexes, through complex processes such as cell division, cell differentiation, germ layer formation and tissue generation, and specific gene expression, the organ system is formed and a complete living individual is formed. After years of development, although people have learned a lot about the phylogenetic process of life, many of its mysteries have not yet been revealed. Among them, the genome sequence and physiological and metabolic conditions of pigs are highly similar to those of humans. Therefore, pigs are considered to be a good model for studying complex human diseases. During the rapid development of pig embryos, the nervous system develops rapidly, and the primordia of the heart, kidney, intestines, and ears also begin to form. Therefore...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/073
Inventor 乔瑞敏李景徐盼吕刚李改英韩雪蕾李新建任广志
Owner HENAN AGRICULTURAL UNIVERSITY
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