RNA-seq of Heterosigma akashiwo: ground control
The TruSeq Stranded mRNA Sample Prep Kit (Illumina, Inc, California, USA) was used. Briefly, total RNA samples (1 ?g) were ribosome-depleted and then reverse-transcribed into double-stranded cDNA with Actinomycin added during first-strand synthesis. The cDNA samples was fragmented, end-repaired and polyadenylated before ligation of TruSeq adapters. The adapters contain the index for multiplexing. Fragments containing TruSeq adapters on both ends were selectively enriched with PCR. The quality and quantity of the enriched libraries were validated using Qubit (1.0) Fluorometer and the Bioanalyzer 2100. The product is a smear with an average fragment size of approximately 360 bp. The libraries were normalized to 10 nM in Tris-Cl 10 mM, pH 8.5 with 0.1% Tween 20.