RNA-seq of Galdieria sulphuraria: hydrolysate replicate 2
Total extraction of RNA was carried out by Monarch Spin RNA Isolation Kit (New England Biolabs, Singapore). RNA quantity and integrity were assessed on an Agilent 2100 Bioanalyzer. Only high-quality RNA sample (OD260/280 = 1.82.2, OD260/230 2.0, RIN6.5, 28S:18S 1.0, >1g) was used to construct sequencing library. Strand-specific mRNA libraries (DNBSEQ Eukaryotic strand-specific) were prepared using poly(A)+ enrichment with oligo-dT beads, mRNA fragmentation, first-strand cDNA synthesis with random primers, second-strand synthesis incorporating dUTP to preserve strandedness, end-repair and 3 adenylation, adaptor ligation, PCR amplification, library QC, library circularization, and DNA nanoball amplification, followed by paired-end sequencing on the DNBSEQ platform.