Arabidopsis thaliana
Protocols: Plants of Arabidopsis thaliana ecotype Col-0 were grown under long day conditions (LD; 16-hours light/8-h dark). Tissue was sectioned into slices with a Leica RM 2155 microtome, distended on pure methanol (Schmid et al., 2012) on Leica PET membrane frameslides, dried at 45 degrees C for 45 minutes, washed for 15 minutes in pure xylol to remove Paraplast, dried again at room temperature for 10 minutes and immediately dissected with a Leica LMD6000 laser microdissector. Material from 3 slides was pooled in a single tube and immediately after microdissection 10 ul of XB buffer from Arcturus Picopure RNA extraction Kit (Life Technologies) was added. Samples were then heated at 42 degrees C for 30 minutes, centrifuged at 800 r.c.f for 2 minutes and stored at -80. Before proceeding with the extraction, material from 5 different tubes were pooled in one; therefore one single replicate contains material microdissected from 15 inflorescences, each harvested from a different plant. RNA extraction was then performed using the Arcturus Picopure RNA extraction kit as specified by the manufacturers instructions. RNA integrity and concentration was tested with an Agilent 2100 Bioanalyzer. Retro-transcription and amplification were performed using the Ovation RNAseq V2 kit (NuGen technologies) as specified by the manufacturers instructions.
ENA-FIRST-PUBLIC
2014-10-02T17:03:58Z
ENA-LAST-UPDATE
2018-03-08T16:47:27Z
External Id
SAMEA2220075
INSDC center name
University of Milan. Dept. of Biosciences
INSDC first public
2014-10-02T17:03:58Z
INSDC last update
2018-03-08T16:47:27Z
INSDC status
public
Submitter Id
E-MTAB-1946:FM_R1
age
1 day
broker name
ArrayExpress
common name
thale cress
developmental stage
meristem stage 1
organism part
flower
sample name
E-MTAB-1946:FM_R1
scientific_name
Arabidopsis thaliana
specimen with known storage state
frozen specimen