RNA-seq to investigate diurnal regulation of gene expression in Medicago truncatula Jester accession.
RNA-seq to investigate diurnal regulation of gene expression in Medicago truncatula Jester accession.
Two frozen trifoliate leaf (per sample) within a 1.5 mL microcentrifuge tube containing two 3.2 mm diameter steel beads (Lab Supply Ltd) was homogenised to a fine powder using the Qiagen TissueLyser II. Tissue was kept frozen using liquid nitrogen. Medicago truncatula Jester accession plants were entrained for 18 days in long days (16h light:8h dark) at 24Â°C constant temperature. Two biological replicates were sampled, per time point every 4h for 24h beginning at ZT0 (dawn) until ZT20 (night) Total RNA was extracted from ground tissue using an RNeasy RNA extraction kit (Qiagen). Extracted total RNA was DNAse-treated (Invitrogen).  Libraries were prepared following the BGISEQ-500 WGS library construction. The full protocol can be found at: https://www.protocols.io/view/bgiseq-500-wgs-library-construction-ps5dng6?step=2