title: ELONGATED HYPOCOTYL 5 (HY5) and POPEYE (PYE) Regulate Intercellular Iron Transport in Plants
design: ELONGATED HYPOCOTYL 5 (HY5) and POPEYE (PYE) Regulate Intercellular Iron Transport in Plants
library_construction_protocol: The roots of seedlings grown on ½ MS for 6 days and transferred to +Fe or −Fe for 3 days were harvested followed by RNA extraction. For seed sterilization, 5% sodium hypochlorite and 70% ethanol was used. After sterilization, seeds were kept at 4°C for 3 days. Seeds were sown on ½ Murashige and Skoog media (Caisson Labs, Smithfield, UT, USA) with Fe (+Fe) or without Fe (-Fe). The media pH was maintained 5.7 with KOH. For making media Fe deficient, 300 μm of [3-(2-pyridyl)-5,6-diphenyl-1,2,4-triazine sulfonate] Ferrozine (Sigma-Aldrich, Bangalore, India) was added in -Fe media. Seedlings were grown under long-day conditions, 16 h light and 8 h dark at 22 °C with a light intensity of 100–110 μmol cm−2 sec−1 and 50% humidity. The soilrite, perlite and compost (3:1:1) mixture was used as soil.  The total RNA was isolated from the roots of seedlings which were grown for 6 days on ½ MS and transferred to +Fe or -Fe for 3 days using the Qiagen Plant RNeasy kit following the manufacturer's protocol. The isolated RNA was treated with DNase I for the removal of genomic DNA. RNA-Seq libraries were prepared using NebNext Ultra II RNA library prep kit for Illumina(E7770L) according to manufacturer's protocols and quality and quantity was assessed using Agilent 4150 TapeStation system and Qubit 4 Fluorometer.
identifiers.namespace: European Bioinformatics Institute
identifiers.id: E-MTAB-14365:PYEHY5T_2_p