title: RNA seq profile of ethylene treated Spirogyra pratensis
design: Total RNA was extracted from Spirogyra filaments using Nucleospin RNA Plant isolation kit (Macherey-Nagel) following the manufacturer’s protocol. RNA quality was assessed using the Experion Electrophoresis System (Bio-Rad) with Experion RNA StdSens chips (Bio-Rad). RNA quantity was measured with a Nanodrop spectrophotometer (Thermo Scientific). Reverse transcription was carried out with the iScript cDNA Synthesis Kit (Bio-Rad) following the manufacturer’s protocol. Purification of mRNA was carried out with the Poly(A) mRNA Magnetic Isolation Module (New England Biolabs) using Agencourt AMPure XP magnetic beads (Beckman Coulter). DNA fragment libraries were prepared for Illumina HiSeq using the NEB Next Ultra RNA Library Prep Kit for Illumina (New England Biolabs) following the manufacturer’s protocol. In order to multiplex the 24 samples on two lanes of the Illumina HiSeq 1500 the Next Multiplex Oligos for Illumina (New England Biolabs) for primer indexing were used.