Detection of MSTN gene mutation
Genomic DNA from transgenic primary cells was extracted using a DNA
extraction kit (Qiagen, Cat. no. 69504). The MSTN primer was
designed using PRIMER3 software
(http://bioinfo.ut.ee/primer3-0.4.0/,
Supplementary Table 1), and the target sequence was amplified by PCR (94
°C for 5 min, 35–40 cycles of 94 °C for 20 sec/57 °C for 30 sec/72 °C
for 35 sec, and 72 °C for 5 min). The PCR product from each sample was
assessed using the T7E1 assay (Toolgene, Cat. no. TGEN_T7E1) to detectMSTN mutations.