title: GSM9269609: N2, EV, D3A, rep1; Caenorhabditis elegans; RNA-Seq
library_construction_protocol: For all experiments, worms were synchronized by bleaching gravid adults to collect eggs, and the resulting synchronized L1 larvae were used to initiate the experiment. All worms were collected at the Day 3 of Adulthood (D3A) stage. Worms were washed multiple times with M9 buffer to remove food, checked under a microscope for purity, and the final pellets were snap-frozen in liquid nitrogen and stored at −80 ∘C prior to RNA extraction. RNA was extracted using RNeasy Kit (Qiagen, #74004). [1] RNA Quality Control:Total RNA was quantified, and its purity was assessed based on OD 260/280 (1.8−2.2) and OD 260/230 (≥1.8) ratios. Only samples with a RIN ≥7.0 were processed for sequencing. 0.1–5 μg of total RNA was used as input. [2] Library Preparation:RNA sequencing libraries were prepared by the Taiwan Genomics Industry Alliance (TGIA) using a standard protocol for Illumina sequencing. The mRNA fraction was enriched via poly-A selection, followed by fragmentation, cDNA synthesis, and adapter ligation. [3] Sequencing:The resulting libraries were pooled and sequenced on an Illumina NovaSeq 6000 System to generate 150 bp, paired−end reads.