title: RNA-seq of Chlorella sp. HS2: sea water at exponential growth phase, biological replicate 1
design: Algal culture from photobioreactor was harvested by centrifugation at 4500 rpm for 10 min. Total RNA was then extracted using the Trizol reagent (Invitrogen, Carlsbad, CA, USA), according to manufacturers instructions. Subsequently, the RNA samples were treated with DNase I for 30 min at 37 C to remove genomic DNA contamination, and the quantity and integrity of the total RNA were verified using an Agilent 2100 bioanalyzer. The cDNA libraries were developed according to manufacturers instructions (Illumina, Inc., San Diego, CA, USA), and sequenced on the Illumina HiSeq 2000 platform at Seeders Co. (Daejeon, Korea) (Liu et al., 2017). In addition, RNA-Seq paired end libraries were prepared using the Illumina TruSeq RNA Sample Preparation Kit v2 (catalog #RS-122-2001, Illumina, San Diego, CA). Starting with total RNA, mRNA was first purified using poly (A) selection, then RNA was chemically fragmented and converted into single-stranded cDNA using random hexamer priming; the second strand was generated next to create double-stranded cDNA. Library construction began with generation of blunt-end cDNA fragments from ds-cDNA. Thereafter, A-base was added to the blunt-end in order to make them ready for ligation of sequencing adapters. After the size selection of ligates, the ligated cDNA fragments which contained adapter sequences were enhanced via PCR using adapter specific primers. The library was quantified with KAPA library quantification kit (Kapa biosystems KK4854) following the manufacturers instructions. Each library was loaded on Illumina Hiseq2000 platform, and the desired average sequencing depth was met while performing high-throughput sequencing.
identifiers.namespace: SUB6550645
identifiers.id: TN1610R1299