Arabidopsis thaliana
Protocols: Five 10-days-old Arabidopsis seedlings were collected into 1,5ml screw cap tubes with 5 glass beads, precooled in liquid nitrogen. Sample homogenizing was done by adding one half of TRI-Reagent (Sigma-Aldrich, 1ml per 100mg) to each sample following sample disruption by using Precellys 24 Lysis & Homogenization instrument for 30sec and 4000rpm. After homogenization total-RNA was extracted by adding 2nd half of TRI-Reagent and the protocol was proceeded according to the manufacture. To remove any co-precipitated DNA, a DNase-I digest was performed by using 1U DNase-I (NEB) in a total Volume of 100ul. Total-RNA was cleaned-up by LiCL-Precipitation using 10µl 8M LiCL and 3Vol 100% Ethanol pa. incubating at -20°C over-night. Following a spin down at 4°C, 13.000rpm for 30min and 2 washing steps with 70% Ethanol pa. The RNA pellet was dried on ice for 1h and resuspended in 40ul DEPC-H2O incubating at 56°C for 5min. The quality of total-RNA was checked by Agilent TapeStation using RNA ScreenTape. Concentration was measured with Qubit RNA BR Assay Kit from Thermo Fisher Scientific.
isolate
not applicable
organism
Arabidopsis thaliana
cultivar
Co-0
age
10
genotype
wild type genotype
developmental stage
seedling
organism part
whole organism
cell_type
plant cell
ENA-FIRST-PUBLIC
2020-10-01
ENA-LAST-UPDATE
2021-08-09