alias: E-MTAB-8158:nrf1-1_-Fe root_s
title: Illumina HiSeq 2000 sequencing; Ribo-Seq analysis of Col-0, nrf1-1 and nrf1-2 mutant under Fe sufficient and Fe deficiency condition
design: Ribo-Seq analysis of Col-0, nrf1-1 and nrf1-2 mutant under Fe sufficient and Fe deficiency condition
library_construction_protocol: 10d old 1/2 MS grown plants transferred to +Fe and -Fe for 2 days and then roots were harvested for polysome isolation. 10d old 1/2 MS grown plants transferred to +Fe and -Fe for 2 days and then roots were harvested for polysome isolation. 10d old 1/2 MS grown plants transferred to +Fe and -Fe for 2 days  mRNA from polysome were extracted with polysome extraction buffer B. Further sucrose gradient was used to isolate the polysome-mRNA complex. Further RNaseI treatment was done to digest the free RNA and ribose boundmRNA was extracted as ribosome protected fragments (RPF). Further purification of RNA was done and send for sequencing. the nucleic acid library was constructed by following the protocol described in Liu et al., 2013, Plant Cell and Ingola et al., 2012, Nature protocol.
identifiers.namespace: Agricultural Biotechnology Research center, Academia Sinica, Taipei 11529, Taiwan
identifiers.id: E-MTAB-8158:nrf1-1_-Fe root_s
attributes.tag: Experimental Factor: genotype
attributes.value: nrf1-1 mutant
attributes.tag: Experimental Factor: growth condition
attributes.value: iron deficiency