Capture and enrichment of genic sequences from a tetraploid individual of the forage grass Festuca ovina, using a sequence capture assay designed in bread wheat
This experiment tested the utility of a bread wheat (Triticum aestivum) sequence capture assay for capturing and enriching genic sequences from the forages species Festuca ovina. The wheat sequence capture array is described in Gardiner et al. 2014 (Plant J, 80, 895-904). It included 110 Mbp of genic sequence information, and is likely to cover the majority of genes present in wheat (Gardiner et al. 2014). Bread wheat is related to Festuca ovina, lying within the same sub-family of the Poaceae (the Pooideae), but in a different tribe. DNA was extracted from leaf tissue of a single individual of Festuca ovina (individual 7015, from microhabitat 1 [deep, high soil pH and moist soil], Bï¿½rby alvar), using a CTAB-phenol-chloroform extraction method with RNase treatment and isopropanol precipitation. DNA quality, quantity and integrity were checked via analysis on the nanodrop platform (ThermoFisher Scientific), via qubit assays (ThermoFisher Scientific), using agarose gel electrophoresis, and on the Agilent Fragment Analyzer platform. Festuca ovina gene-associated sequences were enriched from the DNA sample using the NimbleGen SeqCap EZ in-solution custom capture array designed for bread wheat, using the manufacturers protocol (Gardiner et al. 2014). A paired-end Illumina sequencing library was prepared from the captured DNA fragments. This library was sequenced on one third of a single lane of a HiSeq2000 instrument using 2 x 100 bp paired-end reads.