Affymetrix GeneChip Arabidopsis Genome [AG]

Expression profiling of cytokinin action in Arabidopsis

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10 day old seedlings were treated with 5uM of the cytokinin Benzyladenine(BA)or DMSO at 15min, 45min, 120min, 480min and 1440min

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Transcription profiling by array of Arabidopsis after treatment with benzyladenine

Transcription profiling time series of Arabidopsis seedlings treated with the cytokinin benzyladenine or DMSO

10 day old seedlings were treated with 5uM of the cytokinin Benzyladenine(BA)or DMSO at 15min, 45min, 120min, 480min and 1440min


Affymetrix:Protocol:Hybridization-Unknown

P-AFFY-2

Labeling of samples was carried out according to the manufacturer’s standard protocol

P-AFFY-6

Title: Affymetrix CEL analysis. Description:

P-MEXP-26881

Plants were planted at a density of approximately 500 seeds per plate ( 150 mm) using 0.8% top agar (0.8% low melt agarose in 1X MS salts) on filter paper (Whatman #3, 150mm) placed on 1% agar containing 1X MS salts + 1% sucrose buffered to pH 5.7 with MES (MS media). Seedlings were stratified at 4o C for 4 days, then grown horizontally under constant light (90 uM m-2 s-1) at 22o C for 10 days. The seedlings were then gently scraped off the filter paper into flasks containing 500 ml of liquid MS media and shaken lightly for 2 hr in the light prior to treatment.

P-MEXP-26882

The seedlings were then gently scraped off the filter paper into flasks containing 500 ml of liquid MS media and shaken lightly for 2 hr in the light prior to treatment. At time 0, the indicated concentration of cytokinin (1 or 5 ¥ìM BA or 1 ¥ìM trans-zeatin) was added, or an equal volume of DMSO was added as a vehicle control to the seedlings. After the appropriate time, the seedlings were harvested and frozen at -80¢ª C until RNA was extracted.

P-MEXP-26883

RNA from two independent biological replicates were pooled before hybridizing

P-MEXP-26884

Total RNA was extracted from frozen plant tissue using TRIzol reagent following the manufacture’s instructions (Introgen).

P-MEXP-31507

GeneSpring was used to normalize each chip to a 50th percentile and  the cytokinin treated chips were normalized to the DMSO treated chips. The data that is in the transformed data file is the normalized value and raw value for each Affy probe set on each chip. The DMSO timepoints are self normalized to 1 and are not analyzed further as they are the controls.

Transcription profiling by array of Arabidopsis after treatment with benzyladenine

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