title: Illumina HiSeq 2000 paired end sequencing; Transcriptional responses to cold in five Pooideae species
design: Transcriptional responses to cold in five Pooideae species
library_construction_protocol: Leaf material was collected and flash frozen i) in the afternoon (8 hours after lights on) before cold treatment and the next day 8 hours after cold treatment and ii) in the morning (at lights on) before cold treatment, 4 weeks after cold treatment and 9 weeks after cold treatment. Seeds were germinated and initially grown in a greenhouse at a neutral day length (12 hours of light), 17°C and a minimum artificial light intensity of 150 µmol/m2s. Depending on the species, one to eight week old plants were randomized and distributed to two cold chambers with short day (8 hours of light), 6°C and a light intensity of 50 µmol/m2s. Cold (6°C) with short day (8 hours of light), and a light intensity of 50 µmol/m2s. Flash frozen leaves were individually homogenized using a TissueLyser (Qiagen Retsch) and total RNA was isolated (from each leaf) using RNeasy Plant Mini Kit (Qiagen) following the manufacturer’s instructions. The purity and integrity of total RNA extracts was determined using a NanoDrop 8000 UV-Vis Spectrophotometer (Thermo Scientific) and 2100 Bioanalyzer (Agilent), respectively. Strand-specific TruSeq RNA-seq library prep (Illumina)
attributes.tag: Experimental Factor: Melica nutans
attributes.value: organism
attributes.tag: Experimental Factor: 17
attributes.value: temperature
attributes.tag: Experimental Factor: 12
attributes.value: day length
attributes.tag: Experimental Factor: 150
attributes.value: light intensity
attributes.tag: Experimental Factor: 0
attributes.value: sampling time after light on
attributes.tag: Experimental Factor: control
attributes.value: sampling time point