Plants were stratified for 3 days at 4 degrees in the dark, and then plated on 1/2 B5 + 2% sucrose. Grown at 21 degrees in a 16/8 light/dark cycle conviron chamber for 2 weeks. Plants were transferred to -Fe plates (see Colangelo and Guerinot, Plant Cell 2004) for 72hrs and then roots were harvested and flash frozen in liquid nitrogen.
(Parameters: start time = 2, time unit = weeks, min temperature = 21, temperature unit = C, media = 1/2 B5 + 2%sucrose)
RNA was extracted from plant roots using the standard qiagen plant RNA extraction kit.
(Parameters: Extracted product = total_RNA, Amplification = none)