Ecotype: Columbia Age: Seedling roots, 5 days after germination Growth Media: Standard media for 5 days, transferred to -Fe media for 24 hours before dissection
Treatment protocol
Seedlings were grown for 5 days before transfer to standard media or standard -Fe media. 24 hours after transferring seedlings to media, roots were cut into 4 regions using a razor blade.
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 evenly placed on the mesh in a single row at a density of ~2/cm for all experiments.
Extracted molecule
total RNA
Extraction protocol
Approximately 15 roots were dissected for each replicate and pooled. Samples were collected into RNA extraction buffer and briefly sonicated to disrupt the tissue. 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 longitudinal zone 2 isolated from roots grown under standard conditions for 5 days then transferred to -Fe media for 24 hours
