Fetal skin tissue at 65d, 85d, 105d, 135d, and 7d and 30d after birth (labeled as G1, G2, G3, G4, G5, and G6, respectively) were used, and three samples were collected at each stage. According to the sequences of sheep DNMT1, DNMT3a, DNMT3b, and EDN1 provided by the NCBI, the reference gene was GAPDH, and Primer Premier 5.0 software was used to design the quantitative primers. The cDNA template was used for qRT-PCR amplification. The total system was 25μl, including 1μl cDNA, and each sample had three replicates. The amplification reaction was carried out in Bio-Rad CFX96 fluorescent quantitative PCR instrument.
The gene expressions of WNT2 of the skin, heart, liver, spleen, lung, kidney, and muscle tissue of left shoulder of the lambs at 135d (hair follicle maturation period) were identified. According to the sequences of sheep WNT2, the reference gene was GAPDH (Kang et al., 2020 (link)). The amplification procedures and systems were same as above.
The gene expressions of WNT2 of the skin, heart, liver, spleen, lung, kidney, and muscle tissue of left shoulder of the lambs at 135d (hair follicle maturation period) were identified. According to the sequences of sheep WNT2, the reference gene was GAPDH (Kang et al., 2020 (link)). The amplification procedures and systems were same as above.
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