title: Illumina HiSeq 2500 sequencing; EVOREPRO: RNA-seq of  Marchantia polymorpha Takaragaike sperm cells
library_construction_protocol: To induce sexual reproduction, thalli developed from gemmae were transferred to a far-red light (700 – 780 nm, 44.3 μmol photons m−2 s−1) supplemented light condition using LabLEDs (RHENAC GreenTec Ag).  Male accession of Marchantia polymorpha L., Takaragaike (Tak)-1 was grown on vermiculite under a long-day condition (16/8 h day/night) at 22 °C.  Sperm were released from antheridiophores by applying ddH2O supplemented with RNasin® Ribonuclease Inhibitor (1 u/μl, Promega), collected in a 1.5 mL tube, and pelleted by centrifugation at 3,000 g for 5 min at 4 oC. RNA-seq libraries were generated from total RNA of isolated M. polymorpha sperm using Smart-seq2 (Picelli et al., 2014) using independent biological replicates.