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. Author manuscript; available in PMC: 2023 Jul 21.
Published in final edited form as: Cell. 2022 Jul 21;185(15):2708–2724. doi: 10.1016/j.cell.2022.06.046

Figure 2. Genome Transplantation.

Figure 2.

A. Yeast cells or bacterial cells containing the donor genome to be transplanted are encased in low melt agarose blocks. Yeast cells are spheroplasted using zymolase and digested with proteinase K. B. This leaves the donor genome inside caverns in the agarose, and not sheared during the purification The agarose is melted to gently retrieve the DNA. C. The donor DNA (red) and M. capricolum cells are mixed with polyethylene glycol (PEG) to increase recipient cell membrane fluidity and CaCl2, to mask the DNA charge, resulting in the donor genome entering the recipient cell (at very low frequency). D. The transiently diploid cells are transferred to growth media and begin to grow and divide. E. After several hours, the cells are treated with tetracycline. Only the cells with the synthetic donor genome containing a tetracycline resistance marker survive.