title: Ppa_L1_B_d2
organism: Physcomitrium patens
description: Protocols: The samples were collected every two hours in triplicates with the first sample collected one hour after the onset of the light (ZT1) and the last sample collected 26h later (ZT26) overlapping the first. Cyanophora paradoxa, Physcomitrella patens and Picea abies were grown in 16 h light and 8 h darkness, while Porphyridium purpureum, Klebsormidium nitens, Selaginella moellendorffii were grown in 12 h light and 12 h darkness. Cyanophora paradoxa was grown in C medium at 24°C in 16L/8D photoperiod (40 umol photons m-2 s-1) and areated with normal air.Porphyridium purpureum was grown in ASW medium at 25°C in 12L/12D photoperiod (100 umol photons m-2 s-1).Klebsormidium nitens was grown in C medium at 24°C in 12L/12D photoperiod (40 umol photons m-2 s-1).Selaginella moellendorffii was grown on modified Hoagland solution at 24°C in 12L/12D photoperiod (70 umol photons m-2 s-1). Picea abies was grown on soil at 24°C in 16L/8D photoperiod (250 umol photons m-2 s-1). 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 Selaginella moellendorffii and Physcomitrella patens 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 Cyanophora paradoxa, Porphyridium purpureum and Picea abies were constructed using RS-122-2001/RS-122-2002, TruSeq RNA Library Prep Kit v2 according to the manufacturer's instructions.