GSM9238307: Porcine VIC cultured in osteogenic medium for 8 days, replicate 4; Sus scrofa; RNA-Seq
Total RNA was extracted using the RNeasy RNA extraction kit (Qiagen) following the manufacturer's instructions. RNA integrity was assessed using Agilent 4200 TapeStation (Agilent, Cat# G2991A), and RNA quantity was measured using Qubit 3.0 Fluorometer (Thermo Fisher Scientific, Cat# Q33216) and NanoDrop One spectrophotometer (Thermo Fisher Scientific, ND-ONE). Only RNA samples passing quality control were used for library preparation. mRNA-seq libraries were prepared using the VAHTS Stranded mRNA-seq Library Prep Kit (Vazyme, Cat# NR602) according to the manufacturer's instructions. Library quality and concentration were assessed using Qubit Fluorometer (Thermo Fisher Scientific, Cat# Q33216) and Agilent 4200 TapeStation (Agilent, Cat# G2991A). Sequencing was performed on the Illumina platform. ualified libraries were subjected to Illumina sequencing with a paired-end 150 bp (PE150) strategy. Sequencing was based on the Sequencing by Synthesis (SBS) method: clusters on the flow cell were exposed to four fluorescently labeled dNTPs, DNA polymerase, and sequencing primers. During extension, the incorporation of each labeled dNTP released a specific fluorescent signal, which was captured by the sequencer and converted into sequence information through computational analysis.