Affymetrix GeneChip Arabidopsis Genome [ATH1-121501]

Perception of the Bacterial PAMP EF-Tu by the Receptor EFR Restricts Agrobacterium-Mediated Transformation

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Transcriptional changes upon elicitor treatment over time (0, 30, 60 min) have been analysed with the A.thaliana Landsberg (wt) and fls2-17 (flagellin receptor mutant).

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Transcription profiling of elicitor treatment over time (0, 30, 60 min) in Arabidopsis Landsberg (wt) and fls2-17 (flagellin receptor mutant)

Transcriptional changes upon elicitor treatment over time (0, 30, 60 min) have been analysed with the A.thaliana Landsberg (wt) and fls2-17 (flagellin receptor mutant).


Affymetrix:Protocol:ExpressionStat

Title: Affymetrix CHP Analysis (ExpressionStat). Description:

Affymetrix:Protocol:Hybridization-EukGE-WS2v4

Title: Fluidics Station Protocol. Description:

P-AFFY-6

Title: Affymetrix CEL analysis. Description:

P-MEXP-16928

After a 48-hour treatment at 4 C, A. thaliana Landsberg erecta (Ler-0) and fls2-17 seeds were grown for 12 days on plates containing 1x MS medium (Duchefa), 1% sucrose and 1% agar under continuous light conditions of 60 uE m-2 sec-1 (Biolux lamps) at 22 C. Seedlings were then transferred to liquid MS medium (two seedlings per 500 ul of medium in wells of 24-well-plates). Two days after transfer the medium was supplied with peptides to a final concentration of 10 uM. Plantlets were collected 30/60 min after treatment, frozen in liquid nitrogen and stored at -80 C. 

P-MEXP-16931

Total RNA was prepared using TRIZOL reagent (Life Technologies) and purified on RNeasy Miniprep columns (Qiagen) according to the manufacturers' instructions. 

P-MEXP-16934

After a 48-hour treatment at 4 C, A. thaliana Landsberg erecta (Ler-0) and fls2-17 seeds were grown for 12 days on plates containing 1x MS medium (Duchefa), 1% sucrose and 1% agar under continuous light conditions of 60 uE m-2 sec-1 (Biolux lamps) at 22 C. Seedlings were then transferred to liquid MS medium (two seedlings per 500 ul of medium in wells of 24-well-plates). Two days after transfer the medium was supplied with peptide to a final concentration of 10 uM. Plantlets were collected 30/60 min after treatment, frozen in liquid nitrogen and stored at -80 C. 

P-MEXP-16935

10 ug of total RNA was reverse transcribed using the SuperScript Choice system for cDNA synthesis (Life Technologies) according to the protocol recommended by Affymetrix. The oligonucleotide used for priming was 5'-ggccagtgaattgtaatacgactcactatagggaggcgg-(t)24-3' (Genset Oligo). Double-stranded cDNA was cleaned by phenol:chloroform extraction and the aqueous phase removed by centrifugation through Phase-lock Gel (Eppendorf). In vitro transcription was performed on 1 ug of cDNA using the Enzo BioArray High Yield RNA transcript labelling kit (Enzo Diagnostics). The cRNA was cleaned using RNAeasy clean-up columns (Qiagen). The cRNA was fragmented by heating in 40 mM Tris-acetate pH 8.1, 100 mM KOAc, 30 mM MgOAc. 

Transcription profiling of elicitor treatment over time (0, 30, 60 min) in Arabidopsis Landsberg (wt) and fls2-17 (flagellin receptor mutant)

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