Ecotype: Columbia Age: Seedling roots, 6 days after germination Growth Media: Standard media, transferred to -Fe media for 24 hours prior to harvesting tissue
Treatment protocol
Samples were prepared as in Birnbaum et al. (2005) except that roots were transferred to -Fe media for 24 hours before protoplasting or harvested without protoplasting. Protoplasted cells were passed through the FACS device with all cells being collected.
Growth protocol
Seeds were surface sterilized for 2 minutes in 70% ethanol, the ethanol was removed, then replaced with 30% Bleach and 0.02% Triton X-100 for 15 minutes. Seeds were rinsed 3 times with sterile water, stratified for 4˚C for 2 days, then placed on standard media. Standard media is 1X Murashige and Skoog salt mixture in which ferrous sulfate is replaced with 100mM Fe(III)-EDTA, 0.5g/L MES, 1% sucrose, 1% agar and adjusted to pH 5.7 with KOH. –Fe media is 1X Murashige and Skoog salt mixture in which ferrous sulfate is replaced with 0.3mM Ferrozine. Nylon mesh was placed on top of the solidified media and seeds were planted at ~20 seeds/cm in two rows.
Extracted molecule
total RNA
Extraction protocol
RNA was extracted using the RNAeasy Micro Kit (Qiagen GmbH).
Label
biotin
Label protocol
Fragmented cRNA probes were prepared using the two-cycle amplification protocol by Affymetrix.
Hybridization protocol
Samples were submitted to Expression Analysis Inc. (Durham, NC) for hybridization to Affymetrix ATH1 microarrays.
Scan protocol
Samples were submitted to Expression Analysis Inc. (Durham, NC) for hybridization to Affymetrix ATH1 microarrays.
Description
Gene expression data from whole roots after 24 hours of treatment with -Fe media
