Nanos3_MO_3
Total RNA was extracted using TRIzol reagent and subjected to quality control on an Agilent TapeStation to assess RNA integrity. Samples with sufficient RNA yield were normalized and used for library preparation, which included reverse transcription with barcoded primers, second-strand synthesis, pooling, and clean-up. Amplified RNA (aRNA) was generated by in vitro transcription (IVT) and subsequently fragmented. The aRNA quality was verified by TapeStation electropherogram analysis. Following quality confirmation, the aRNA was reverse-transcribed to complementary DNA (cDNA) and amplified by PCR. The resulting cDNA libraries were checked on the TapeStation for fragment size distribution, purified, and submitted for high-throughput sequencing.