Method for promoting cell to differentiate into female genital cell based on overexpression of Figla gene
A germ cell and gene technology, applied in the field of promoting cell differentiation to female germ cells based on the overexpression of Figla gene, can solve the problems of limited number of differentiation, long induction process, poor induction efficiency and repeatability
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[0038] The present invention will be further described in detail below in conjunction with specific embodiments, which are for explanation rather than limitation of the present invention.
[0039] 1. Figla gene cloning and vector construction
[0040] According to the gene sequence of mouse Figla in the GenBank database (NCBI: NM_012013.1), Primer Premier 5.0 primer design software was used to design and amplify the upper and downstream primers of Figla gene and mutate the stop codon of Figla gene, and select the appropriate restriction endonuclease Enzyme, specifically design the following primers:
[0041] Upstream primer: 5'TC AGATCT TGGTCTTGACCACCATGGATA 3′
[0042] Downstream primer: 5′CT GAATTC CATCAGACTCCTCATGAGTGAAGTA 3′
[0043] The underlined part in the upstream primer is the Bgl II restriction site, and the underlined part in the downstream primer is the EcoI restriction site;
[0044] Figla gene PCR amplification reaction system is 15μL, including: 10×buffer 1.5μL, MgCl2 ...
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