GSM1347648: HC_12h replicate2; Nannochloropsis oceanica; RNA-Seq
Algal total RNAs under the above conditions were extracted using Trizol reagents (Invitrogen). For RNA-Seq, the poly (A) containing mRNA molecules were purified using Sera-mag Magnetic Oligo(dT) Beads (Thermo Scientific) and was fragmented into 200- to 300-bp fragments by incubation in RNA Fragmentation Reagent (Ambion)according to the manufacturer’s instructions. The fragmented mRNA was then purified away from the fragmentation buffer by Agencourt®RNAClean beads (Beckman Coulter). The purified, fragmented mRNA was then converted into double-stranded cDNA using the SuperScript Double-Stranded cDNA Synthesis Kit (Invitrogen) by priming with random hexamers. Strand nonspecific transcriptome libraries were prepared using NEBNext® mRNA Library Prep Reagent Set(New England Biolabs) and sequenced for 2×90bprun(paired-end, PE) using Illumina HiSeq2000.
GEO Accession
GSM1347648