title: GSM7030450: AC-24h-endo-rep1; Avena sativa; RNA-Seq
library_construction_protocol: The RNA was extracted using Spectrum™ Plant Total RNA Kit from Sigma-Aldrich (ref. STRN250-1KT) with some additional optimisation steps. Samples were pre-treated with 0.03% (v/v) α-amylase (Sigma-Aldrich, ref. A4551-100MG) in the lysis buffer. Ground frozen samples were vortexed for 1 min with the mixture and incubated at RT for 6 min. 0.01 % (v/v) β-mercaptoethanol was added to the samples and vortexed again for 30 sec. Samples were centrifuged at RT for 5 min at max r.p.m. Supernatant was transferred to the filtering columns and Spectrum™ Plant Total RNA protocol was used following manufacturer's instructions including the on-column DNase treatment. Protocol A was used for the binding buffer solution. Samples were further cleaned up using RNA Clean & Concentrator-5 kit (Zymo Research, ref. R1016) to ensure purity and concentration. Libraries were generated using TruSeq® Stranded mRNA Library Prep Kit (Illumina) according to manufacturer's instructions and sequenced on a Nextseq 500 platform (Illumina) as single-end reads.