Animal preparation method

A method for preparing a non-human animal, comprising aggregating a tetraploid embryo with embryonic stem cells to form a new reconstructed embryo or a chimeric embryo, wherein the tetraploid embryo is a tetraploid embryo at 2-cell stage;

• preferably, the method adopts a tetraploid complementation assay or a tetraploid embryo complementation assay;
• preferably, the method comprises the following steps:
• (1) obtaining an animal 2-cell embryo;
• (2) placing the 2-cell embryo obtained in step (1) in a fusion solution and performing fusion to obtain a tetraploid embryo;
• (3) placing the tetraploid embryo obtained in step (2) in a culture medium and culturing;
• (4) aggregating the tetraploid embryo, which is developed to 2-cell stage in step (3), with embryonic stem cells to form a chimeric embryo;
• preferably, in step (3), the tetraploid embryo is cultured for about 8-24 hours;
• or preferably, the culture medium in step (3) is KSOM medium.

A set of primers, comprising:

• an upstream primer that has at least 85% or at least 90% or at least 91% or at least 92% or at least 93% or at least 94% or at least 95% or at least 96% or at least 97% or at least 98% or at least 99%, or 100% identity to a sequence as shown by SEQ ID NO:
• 19;
• and a downstream primer that has at least 85% or at least 90% or at least 91% or at least 92% or at least 93% or at least 94% or at least 95% or at least 96% or at least 97% or at least 98% or at least 99%, or 100% identity to a sequence as shown by SEQ ID NO:
• 20.