title: RNA-seq on single seedlings for 24 seedlings of Col0-WT grown with low nitrate on petri dishes
library_construction_protocol: 11-day-old seedlings were harvested individually in liquid nitrogen Col0 WT plants were grown on petri dishes after seed sterilisation and stratification, on a solid media containing 1mM KH2PO4, 1mM MgSO4, 0.25mM K2SO4, 0.25mM CaCl2, 0.1mM Na-Fe-EDTA, 2.5mM MES and microelements (50μM KCl, 30μM H3BO3, 5μM MnSO4, 1μM ZnSO4, 1μM CuSO4 and 0.1μM (NH4)6Mo7O24) and 1mM KNO3. A total of 4 plates per genotype and condition were prepared, with 30 seeds per plate (disposed over 2 rows). After 6 days of growth on long days at 22 degrees Celsius and a light intensity of 150 μmol m-2 s-1, seedlings that managed to do the heterotrophic-to-autotrophic transition and continued to grow were transferred to a new plate. RNA was extracted from 11-day-old individual seedlings using the MagMAX™-96 Total RNA Isolation Kit (Ambion) following manufacturer's instructions except for two changes: 60µl of Lysis/Binding solution were used instead of 100µl, and the elution was performed with 30µl of Elution Buffer instead of 50µl.  Library for sequencing was performed by the novogene company on total RNA using a polyA enrichment method. 