STRT-seq of Sus scrofa: wild-type pEPSCs
Single-cell libraries were prepared using a modified STRT-seq method as previously described (Dong et al., 2018). Following digestion, single pEPSC cells were randomly selected using a mouth pipette and transferred into lysis buffer containing an 8-bp barcode. First-strand cDNA synthesis and amplification were conducted in a reverse transcription (RT) mixture comprising 4 U RNase inhibitor, 100 U SuperScript II reverse transcriptase (Invitrogen, 18064071), 1 mM dNTPs (TAKARA, 4019), 60 mM MgCl2, and 3 M RT primer along with 10 M TSO primer. The amplified product underwent purification twice with 0.8 AMPure XP beads (Beckman, A63882). Subsequently, biotin-based PCR enrichment was performed. The scRNA-seq library was constructed using the KAPA Hyper Prep Kit (KAPA, KK8054) following the manufacturers protocol for PCR Library Amplification/Illumina platforms. High-quality libraries were sequenced with 150-bp paired-end reads on an Illumina NovaSeq 6000.