Long Non-Coding RNA Sequencing from Chicken Embryo

Total RNA extraction from chicken embryo visceral tissues (CEVTs) without any modification was previously described in detail in our study [12 (link)]. Thereafter, the qualified RNA samples for library construction underwent tRNA and rRNA removal treatment before cDNA library preparation to meet the specified requirements. The preliminary cDNA library was further processed by performing cDNA end repair and Ploy A tail ligation—an essential component of lncRNA—followed by PCR reaction to obtain a complete cDNA library. This was performed according to the manufacturer’s instructions, without any modification (Illumina Stranded Total RNA Prep with Ribo-Zero Plus kit, Illumina, San Diego, CA, USA). After the purification and quality assessment of the constructed libraries, lncRNA sequencing was conducted on an Illumina NextSeq 2000 Sequencing System (Illumina, San Diego, CA, USA).