|
Status |
Public on Jan 11, 2011 |
Title |
apx1/cat2 plants-1h vs 0h HL-Rep2 |
Sample type |
RNA |
|
|
Channel 1 |
Source name |
apx1/cat2, 0h HL
|
Organism |
Arabidopsis thaliana |
Characteristics |
genotype: apx1/cat2 treatment: none time: 0 hour plant age: 32 days
|
Treatment protocol |
3- to 4-week-old plants (at the same developmental stage of 1.09-1.10, Boyes et al., 2001) were exposed to a continuous high light irradiation (1000 μmol.m-2.s-1) in a SANYO growth cabinet for 0 or 1 hour.
|
Growth protocol |
Arabidopsis thaliana plants were grown on Jiffy-7 soil pellets (Jiffy Products, Norway) in a controlled growth chamber (Weiss technik) at 21-22 °C, 16 h/8 h light regime at 100 μmol.m-2 s-1 photoperiod and 70% relative humidity.
|
Extracted molecule |
total RNA |
Extraction protocol |
From two independent experiments, RNA was isolated from middle-aged leaves of 25 wild type (WT), apx1, cat2, and apx1/cat2 plants using TRIzol Reagent (Invitrogen, Carlsbad, CA). Concentration and quality of the RNA was determined with a Nanodrop ND 1000 spectrophotometer and quality was examined using the RNA 6000 Nano Assay (Agilent Technologies 2100 Bioanalyzer).
|
Label |
Cy5
|
Label protocol |
Per sample, an amount of 1 µg of total RNA was converted to double-stranded cDNA in a reverse transcription reaction. Subsequently, the sample was converted to antisense cRNA, amplified and labeled with Cyanine 3-CTP (Cy3) or Cyanine 5-CTP (Cy5) in an in vitro transcription reaction according to the manufacturer’s protocol (Agilent).
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|
|
Channel 2 |
Source name |
apx1/cat2, 1h HL
|
Organism |
Arabidopsis thaliana |
Characteristics |
genotype: apx1/cat2 treatment: high light irradiation time: 1 hour plant age: 32 days
|
Treatment protocol |
3- to 4-week-old plants (at the same developmental stage of 1.09-1.10, Boyes et al., 2001) were exposed to a continuous high light irradiation (1000 μmol.m-2.s-1) in a SANYO growth cabinet for 0 or 1 hour.
|
Growth protocol |
Arabidopsis thaliana plants were grown on Jiffy-7 soil pellets (Jiffy Products, Norway) in a controlled growth chamber (Weiss technik) at 21-22 °C, 16 h/8 h light regime at 100 μmol.m-2 s-1 photoperiod and 70% relative humidity.
|
Extracted molecule |
total RNA |
Extraction protocol |
From two independent experiments, RNA was isolated from middle-aged leaves of 25 wild type (WT), apx1, cat2, and apx1/cat2 plants using TRIzol Reagent (Invitrogen, Carlsbad, CA). Concentration and quality of the RNA was determined with a Nanodrop ND 1000 spectrophotometer and quality was examined using the RNA 6000 Nano Assay (Agilent Technologies 2100 Bioanalyzer).
|
Label |
Cy3
|
Label protocol |
Per sample, an amount of 1 µg of total RNA was converted to double-stranded cDNA in a reverse transcription reaction. Subsequently, the sample was converted to antisense cRNA, amplified and labeled with Cyanine 3-CTP (Cy3) or Cyanine 5-CTP (Cy5) in an in vitro transcription reaction according to the manufacturer’s protocol (Agilent).
|
|
|
|
Hybridization protocol |
A mixture of purified and labeled cRNA was hybridised on Agilent arrays followed by (manual) washing, according to the manufacturer’s procedures.
|
Scan protocol |
Arrays were scanned using the Agilent DNA MicroArray Scanner. Images were quantified using Agilent Feature Extraction Software (version 8.1.1.1).
|
Description |
Biological replicate 2 of 2. apx1/cat2 plants subjected to high light, harvested after 0h and 1h.
|
Data processing |
Features were extracted with spatial detrending and dye normalization (Linear Lowess) using the Agilent Feature Extraction Software (version 8.1.1.1).
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|
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Submission date |
Jan 12, 2010 |
Last update date |
Jan 11, 2011 |
Contact name |
Rekin's Janky |
E-mail(s) |
Nucleomics.Bioinformatics@vib.be
|
Organization name |
VIB
|
Department |
Nucleomics Core
|
Street address |
Herestraat 49 Box 816
|
City |
Leuven |
ZIP/Postal code |
B-3000 |
Country |
Belgium |
|
|
Platform ID |
GPL2871 |
Series (1) |
GSE19857 |
Functional characterization of Arabidopsis mutants deficient in cytosolic APX1 and peroxisomal CAT2 |
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