Illumina HiSeq 2500 sequencing; Comparative analysis of abiotic stress responses in the basal algae Cyanophora paradoxa
Comparative analysis of abiotic stress responses in the basal algae Cyanophora paradoxa
The samples were harvested by centrifugation (4000 rpm, 2 min)  Fresh cultures were grown for 7 days before the stress was applied in C medium at 24°C in continuous light (40 umol photons m-2 s-1) and aeration with ordinary air. For the nutrient stresses the cultures were washed twice (300g, 10 min, 4°C) with new media and then transferred to the growth chamber (24°C, continuous light 40 umol photons m-2 s-1, aeration with ordinary air). For the environmental stresses the cultures were grown without aeration and subjected to the stress for 12h. The prolonged darkness cultures were placed in the dark at 24°C  60h before. Cold, heat and control were grown in the respective conditions in the dark. The total RNA was extracted using Spectrum™ Plant Total RNA Kit (Sigma-Aldrich) according to the manufacturer's instructions. The libraries were prepared from total RNA using polyA enrichment. The cDNA libraries for the environmental stresses samples were constructed using the NEBNext Ultra™ Directional RNA Library Prep Kit for Illumina (New England Biolabs) according to the manufacturer's instructions using 500 ng of Input material with 10 min fragmentation time and 15 PCR cycles. The cDNA libraries for the nutrient stresses samples were constructed using RS-122-2001/RS-122-2002, TruSeq RNA Library Prep Kit v2 according to the manufacturer's instructions.
Experimental Factor: stimulus
environmental stress
Experimental Factor: growth condition
cold (4C), in darkness for 12h