GSM9360748: Control-1 Treatment; Coelastrella tenuitheca; RNA-Seq
Total RNA was extracted from 50 mg of algal biomass using TRIzol reagent following the manufacturer's protocol, with DNase I treatment to remove genomic DNA contamination. RNA quality was assessed using an Agilent Bioanalyzer, and only samples with RIN > 8.0 were used for library construction. RNA-seq libraries were prepared from 1 μg of total RNA using the NEBNext Ultra II Directional RNA Library Prep Kit. Poly(A) mRNA was selected using oligo-dT magnetic beads, followed by fragmentation and first-strand cDNA synthesis. Second-strand cDNA was synthesized with dUTP for strand specificity. After end repair, A-tailing, and adapter ligation, libraries were size-selected (300-500 bp) and PCR-amplified for 12 cycles. Final libraries were quantified by Qubit and quality-checked by Bioanalyzer.