① Prepare embryos at the blastula stage, surgical needles, and operating needles.
② Place one culture medium in an agarose petri dish and place the embryo to be operated on.
③ The yolk part of one embryo (donor) is held by an operating needle, and the blastoderm is cut along the latitude line with a surgical needle to form a blastoderm fragment (graft).
④ Similarly, while holding down the yolk portion, a cut is placed in the transplanted portion of the other embryo (host).
⑤ The donor embryonic cleavage site overlaps the host cleavage site. It is necessary to do it within one minute after it is first cut until it is piled up.
⑥ After 2-3 minutes, hold the donor and the cut surface of the host so that they stick to each other again.
⑦ After confirming that the grafted portion is bound, culture one by one in a 96-well culture plate filled with the primary culture medium containing antibiotics. At this time, the embryo or yolk fragment contained in the culture medium at the time of operation should not be placed in the well of the culture plate.
