Fig. 2

Sequence analysis of the MSTN locus in transfected cell strains. a Surveyor nuclease cleavage of the transfected cell strains. The amplicons from 21 cell strains were digested by restriction enzyme Alu I. Two bands were observed in wild-type cell strains, as shown in lanes 12, 14, and 15. Three bands were observed in mono-allelically modified cell strains or mixture cells, containing a 444-bp additional band, as shown in lanes 1–3, 5, 8, 9, 11, and 19–21. Only one 444-bp band was observed in bi-allelically modified cell strains, as shown in lanes 4, 6, 7, 10, 13, and 16–18. b Sequence variations at the cleavage site of 10 donor cell strains were aligned with the wild-type sequence. L52, L383, and L385 had mono-allelic mutations; L95, L170, L179, and L231 had bi-allelic mutations; and L193, L301, and L323 had homozygous mutations. ‘-’ means frameshift mutant; ‘+’ means insertion mutant. c Sequence analysis of the cloned fetuses. Three cloned fetuses (named C231-1, C231-2, and C323) were obtained from two recipients. C231-1 and C231-2, containing the same genotype, came from the bi-allelic mutant donor cell L231. C323 came from the homozygous mutant donor cell L323