Paspalum dilatatum transcriptome
Plant tissues were harvested for RNA isolation from a single clone of P. dilatatum cv. Primo at the late reproductive stage, from leaves, stems, roots and mature inflorescences. Total RNA was isolated using RNeasy* Plant Mini Kit (Qiagen, Hilden, Germany). A total of 250 ng of total RNA from each RNA extraction was combined to provide a total of 1 *g for use in cDNA synthesis using the SMART™ cDNA Synthesis kit (Clontech Laboratories, Mountain View, USA), a modified poly-T primer sequence (5’-AAGCAGTGGTATCAACGCAGAGTCGCAGTCGGTACTTTTTTCTTTTTTV-3’) was used instead of the CDS-II primer to reduce sequencing problems associated with the poly-A tails. Approximately 5 *g of the amplified cDNA library was sheared by nebulisation at 200 kPa for 2 min. The GS-FLX titanium shotgun sequence library was then prepared following the manufacturer’s instructions (Roche Diagnostics, Basel, Switzerland) and enriched beads were loaded onto one half of a picotitre plate for sequencing.