Instrument: Illumina HiSeq 2500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Main shoot apex tissue was isolated at Waddington stage W3.5 and 5.5 and frozen in liquid nitrogen. Tissue was ground, dissolved in 500 mL TRIzol reagent (Invitrogen), incubated for 5 min at room temperature after which, 100 mL of chloroform was added. The sample was homogenized, incubated for 2 min, and centrifuged for 15 min at 4°C followed by phase separation (keeping the aqueous phase). Next, isopropanol was added and incubated for 10 min at room temperature and further purified using an RNA easy Micro Kit (Qiagen). Residual DNA was removed using a DNA-free kit (Ambion). The quality of the RNA was tested using a bio analyzer (Agilent). RNA libraries were prepared for sequencing using standard Illumina protocols