RNA-seq of 77S stem replicate 2
RNA sequencing libraries were constructed using the Illumina TruSeq Stranded mRNA Library Prep Kit following the manufacturer's instructions. Briefly, 500 ng of total RNA was purified using poly-T oligo-attached magnetic beads to select for poly(A)-containing mRNA. The purified mRNA was then fragmented, and first-strand cDNA was synthesized using random hexamers and reverse transcriptase, followed by second-strand cDNA synthesis. The double-stranded cDNA underwent end-repair, A-tailing, and adapter ligation. The final library was enriched by PCR and purified using AMPure XP beads. The quality and concentration of the library were assessed using an Agilent 2200 TapeStation.