title: RNA-seq of Desmodesmus sp. G41 mutant
design: Total RNAs were extracted using the TRIzol reagent and were then treated with RNAase-free DNase I (TaKaRa, Japan) to remove any contaminating genomic DNA. RNA purity was checked using the NanoPhotometer spectrophotometer (IMPLEN, CA, USA), and RNA integrity was assessed using the RNA Nano 6000 Assay Kit of the Agilent Bioanalyzer 2100 system (Agilent Technologies, CA, USA).  A total amount of 1.5 g RNA per sample was used to generate mRNA-seq libraries of Desmodesmus. sp using NEBNext Ultra RNA Library Prep Kit for Illumina (NEB, USA) according to manufacturers recommendations. Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads and then fragmented. Double-stranded cDNAs were synthesized using random hexamer primer and M-MuLV Reverse Transcriptase(RNase H-). Then mRNA-seq libraries were constructed following the standard Illumina protocol of adapter ligation with NEBNext Adaptor and PCR amplification with Phusion High-Fidelity DNA polymerase. PCR products were purified by AMPure XP system and library quality was assessed on the Agilent Bioanalyzer 2100 system, to generated 150-200bp paired-end fragments. Subsequently, an Illumina HiSeq4000 platform was applied for the library sequencing .