Setaria italica maturation reads
Total RNA was extracted from 100mg of tissue using the Bio-Rad Aurum Total RNA kit for fatty and fibrous tissue as per the manufacturer’s instructions, including the on column DNaseI treatment. The quality of the RNA was determined using the Agilent Bioanalyser before proceeding (RIN = 7.6 – 8.7; data not shown). RNA sequencing, including library preparation, was performed by the genomics facility at the Kinghorn Centre for Clinical Genomics (Garvan Institute of Medical Research, 384 Victoria St, Darlinghurst, NSW 2010, Australia). Briefly, RNA-Seq libraries were prepared using the Illumina TruSeq Stranded mRNA Library kit using the standard protocol to produce libraries with an average size of 309bp. These libraries were then sequenced on an Illumina HiSeq 2500 system (HiSEQ Control Software 2.2.38, RTA 1.18.61) following the standard rapid sequencing workflow. Samples were loaded at a concentration of 14pM and a total of 209 cycles of sequencing were completed consisting of 2 101bp reads and a single 7bp index sequence.