title: RNA-seq of Z. muelleri: leaf tissue
design: Sterile, pre-chilled mortar and pestles were used to grind leaf tissue into fine powder in liquid nitrogen. RNA was extracted from 80 130 mg of fresh weight leaf tissue using an Ambion PureLink RNA Mini kit (Fisher Scientific) and an Ambion PureLink DNase Set (Thermo Fisher Scientific). The quantity and quality of the extracted RNA was measured using a 2100 Bioanalyzer Instrument (Agilent Technologies). All RIN (RNA Integrity) numbers were above 7, indicating high quality material was used for sequencing. mRNA was isolated from total RNA using a TruSeq mRNA Stranded Total Library Prep Kit (Illumina) and then sequenced using the HiSeq2500 System (Illumina) at The Australian Cancer Research Foundation (ACRF) Biomolecular Resource Facility (BRF), Australian National University.