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A cloned embryo treatment liquid, its use method and the application of the treatment liquid

A technology for processing liquids and embryos, applied in embryonic cells, biochemical equipment and methods, cells modified by introducing foreign genetic material, etc., and can solve the problem of low efficiency

Active Publication Date: 2021-06-25
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Because of this advantage, the use of ESc for somatic cell cloning often has a high cloning efficiency, but its efficiency is still not high

Method used

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  • A cloned embryo treatment liquid, its use method and the application of the treatment liquid
  • A cloned embryo treatment liquid, its use method and the application of the treatment liquid
  • A cloned embryo treatment liquid, its use method and the application of the treatment liquid

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Experimental program
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Embodiment Construction

[0029] The present invention will be described in further detail below in conjunction with the accompanying drawings.

[0030] Experimental Materials

[0031] 1. Cell lines and pig cloned embryos

[0032] Pig adult fibroblasts (513D) were collected from breeding pigs provided by Guangdong Wen's Breeding Pig Technology Co., Ltd. Shuitai Original Pig Farm; pig cloned embryos were prepared in the cloning laboratory of Guangdong Wen's Breeding Pig Technology Co., Ltd.

[0033] 2. Main reagents and consumables

[0034]RNA extraction kit E.Z.N.A.Total RNA Kit I was purchased from OMEGA Company; embryo RNA extraction kit Qiagen RNeasy Plus Micro Kit (50) was purchased from Qiagen Company; reverse transcription kit PrimeScriptTMRTreagent Kit with gDNA Eraser was purchased from TaKaRa Company; Quantitative PCR kit SYBR SelectMaster Mix was purchased from Life Technologies; Taq enzyme for general PCR was purchased from Guangzhou Dongsheng Biotechnology Company; DL500 DNA Marker was pu...

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Abstract

The invention discloses a cloned embryo treatment solution, which is composed of BIX-01294 and PZM culture solution, the concentration of BIX-01294 in the PZM culture solution is 5nM-500nM; wherein, the PZM culture solution is composed of the following components: NaCl 3.156g; NaHCO 3 1.053g; KCl 0.373g; KH 2 SO 4 0.024g; MgSO 4 0.049g; calcium lactate 0.308g; pyruvate 0.011g; L-glutamic acid 0.073g; hypotaurine 0.273g; BME 10mL; MEM 5mL; penicillin 0.033g; 500mL. Using the cloned embryo treatment solution of the above technical scheme to culture and treat cloned embryos can achieve the following beneficial effects: (1) can significantly reduce the gene expression of cells SUV39H1, SETdb1, DNMT1 or DNMT3a; (2) can significantly down-regulate the H3K9me2 levels in 2cell and 4cell.

Description

technical field [0001] The invention relates to somatic cell cloning technology, in particular to a treatment solution for cloning embryos, a method for using the treatment solution and the use of the treatment solution. Background technique [0002] Somatic cell nuclear transfer (SCNT), also known as somatic cell cloning, is an assisted reproductive technology used to produce animals. SCNT refers to the transplantation of donor somatic cells into the cytoplasm of enucleated oocytes, through fusion and activation to obtain reconstructed embryos, allowing the cells to resume division and continue to develop to obtain offspring with the same genotype as the donor cells. [0003] Through somatic cell cloning technology, the nucleus of a single differentiated somatic cell can be cultured in an in vitro environment to obtain a whole biological organism, which has great potential for agricultural technology, biomedicine industry, and protection of endangered species (Yang et al., ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/10C12N5/075
CPCC12N5/0604C12N5/0609C12N2500/05C12N2500/30C12N2500/32C12N2500/33
Inventor 吴珍芳杨林雪杨洋赵成法贺晓燕蔡更元李紫聪刘德武
Owner SOUTH CHINA AGRI UNIV
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