T1F1S1
Zea mays
Protocols: Suitable farms were selected by KALRO extension officers, spaced at least 10km apart, with emphasis placed on getting samples from a wide geographic area. Leaf tissue was detached using clean tweezers, submerged in RNA later (Ambion) and stored on dry ice until a -40 freezer was reached. Sampled maize plants were grown outside, and infected naturally in farmers fields Symptomatic plants were selected from farmers fields under natural infection conditions RNA was extracted using Trizol (Ambion). RNA treated using Ribo-Zero Magnetic Kit (Plant Leaf-Epicentre). Indexed stranded libraries were constructed using Scriptseq V2 RNA-Seq Library Preparation kits and Scriptseq Index PCR primers (Epicentre). Purification steps were performed using Agencourt AMPure XP beads. Library quantity and quality were checked using Qubit (Life Technologies) and on a Bioanalyzer High Sensitivity DNA Chip (Agilent Technologies).