Illumina NovaSeq 6000 paired end sequencing; RNA-seq of dinoflagellate Heterocapsa sp. strain CCAP 1125/4
RNA-seq of dinoflagellate Heterocapsa sp. strain CCAP 1125/4
Sample for bulk RNA was collected by filtering 10 ml of culture in exponential phase (density of cells ≈ 10 mil) on 5 µm polycarbonate filters (Whatman).Filters were placed in cryoprotectant tubes with beads (Zirconium oxide beads, Precellys), amended with 600 µl of RLT buffer (Qiagen) which contained 1% of 2-Mercaptoethanol, shock frozen in liquid nitrogen and stored at -80 °C until further processing. Prior to RNA extraction, sample was taken out of -80, thawed on ice and subjected to bead beating (1x 6000 rmp, Precellys Evolution homogenizer)for 2 min. RNA from samples was further extracted with the RNeasy Qiagen kit (QIAGEN, Inc., Valencia, CA) following the kit instructions. RNA was re-eluted in 30 µl of nuclease free water. Input: RNA Type: Illumina TruSeq Stranded mRNA Option: Poly-A selection Category: default