GSM7445450: 1h_nFe3O4 (2c); Raphidocelis subcapitata; RNA-Seq
RNA isolation of microalgae R. subcapitata involved two major steps: the cell lysis and then the RNA extraction procedure. RNA was harvested from three biological replicates after 1 and 4 h from R. subcapitata exposed with nZVI and nFe3O4 respectively. Cell pellets were obtained through centrifugation at 10,000 × g, 4°C for 2 min, followed by overnight cell lyophilization and phenol-chloroform extraction method adapted from a previous work (Yeap et al., 2021). The integrity and concentration of the RNA extracts were determined with an Agilent 2100 Bioanalyzer system (Agilent Technologies, Germany) and Nanodrop (Thermo Scientific, USA) was for RNA concentration quantification. All electropherogram of ribosomal RNA peaks were computed and analyzed by 2100 Expert software (Agilent Technologies, Germany). To prepare for whole transcriptome sequencing of the directional RNA-seq library, up to 1 µg of total RNA (RIN > 6) was used with a TruSeq® stranded mRNA Library Prep kit (Illumina Inc., USA) after capturing poly-adenylated transcripts and mRNA fragmentation. The resulting cDNA libraries were pooled and sequenced in paired-end mode (2 × 79 bp) on an Illumina NextSeq 500/550 High Output v2 platform.