title: Illumina HiSeq 2500 paired end sequencing; Effect of long-term high CO2 environment on gene expression of Phaeodactylum tricornutum
design: Effect of long-term high CO2 environment on gene expression of Phaeodactylum tricornutum
library_construction_protocol: P.tricornutum cells from three biological replicates were harvested in the middle of light period by vacuum filtration on 1.2 um polycarbonate filters (Millipore, United States) long time(8 months).  P. tricornutum cells were harvested in the middle of the light period by vacuum filtration onto 1.2 um polycarbonate filters (Millipore, United States), then flash frozen in liquid N2 and maintained at -80C until RNA extraction. The LC sample comprised three biological replicates and HC sample comprised three biological replicates. P.tricornutum cells from three biological replicates were harvested in the middle of light period by vacuum filtration on 1.2 um polycarbonate filters (Millipore, United States) long time(8 months).  P. tricornutum cells were harvested in the middle of the light period by vacuum filtration onto 1.2 um polycarbonate filters (Millipore, United States), then flash frozen in liquid N2 and maintained at -80C until RNA extraction. The LC sample comprised three biological replicates and HC sample comprised three biological replicates. Total RNA was extracted from the frozen cell pellets using the TRIzol Reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's instructions. Total RNA was extracted using the mirVana miRNA Isolation Kit (Ambion) following the manufacturer's protocol. RNA integrity was evaluated using the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA). The samples with RNA Integrity Number (RIN) 7 or higher were subjected to the subsequent analysis. The libraries were constructed using TruSeq Stranded mRNA LTSample Prep Kit (Illumina, San Diego, CA, USA) according to the manufacturer's instructions.