Protocols: For each individual plant (e.g. segregating wild type or RNAi line) a pooled sample of four mature leaves was collected and flash frozen in liquid nitrogen. The frozen tissue was ground using of a Tissuelyser (Qiagen TissueLyser II) while keeping the samples frozen. Plants were grown in a greenhouse in Oxford. The frozen tissue was suspended in 1 ml TRIzol reagent (Thermo Fisher) reagent using a vortex until the powder was fully suspended. Subsequently, the sample was incubated at room temperature for 5 min. Next, the cell debris were collected through centrifugation at 15'000 g at 4°C for 10 min. The supernatant was transferred to a 1.5 ml Eppendorf tube containing 200 μl chloroform and thoroughly mixed. The solution was centrifuged at 15'000 g at 4°C for 15 min and the upper aqueous phase was transferred to a fresh 1.5 ml Eppendorf tube containing 500 μl isopropanol. The solution was mixed and kept at room temperature for 10 min. Next, the extracted RNA was collected by centrifugation at 15'000 g at 4°C for 10 min. The supernatant was poured off and the RNA pellet was washed through resuspension in 75% ethanol and collected using a centrifuge at 15'000 g at 4°C for 5 min. The supernatant was removed and the wash step was repeated. Next, the RNA pellet was resuspended in 50 μl water (diethylpyrocarbonate treated) and the remaining DNA was removed using TurboDNAse (Thermo Fisher) treatment. The RNA was shipped to BGI for single-end transcriptome sequencing. Single end Illumina protocol
ENA first public
2022-01-01
ENA last update
2022-01-01
External Id
SAMEA6651318
INSDC center alias
OXFORD
INSDC center name
University of Oxford
INSDC first public
2022-01-01T00:12:21Z
INSDC last update
2022-01-01T00:12:21Z
INSDC status
public
Submitter Id
E-MTAB-8907:segWT_31
broker name
ArrayExpress
common name
foxtail millet
cultivar
Setaria viridis
genotype
wild type genotype
organism part
leaf
sample name
E-MTAB-8907:segWT_31