comparative transcriptome sequencing in the marine dinoflagellate Prorocentrum lima under phosphorus limitation
Sequencing libraries were generated using NEBNext UltraTM RNA Library Prep Kit for Illumina (NEB, USA) following manufacturers recommendations as described in previous paper (Wan et al., 2023). Briefly, mRNA was purified from total RNA using poly-T oligo-attached magnetic beads. First strand cDNA was synthesized using random hexamer primer and M-MuLV Reverse. Second strand cDNA synthesis was subsequently performed using DNA polymerase I and RNase H. In order to select cDNA fragments of preferentially 250~ 300 bp in length, the library fragments were purified with AMPure XP system (Beckman Coulter, Beverly, USA), and library quality was assessed on the Agilent Bioanalyzer 2100 system. After cluster generation, the library preparations were sequenced on an Illumina Novaseq 6000 platform and 150 bp paired-end reads were generated.