Exp1_leaf_Aonly_rep2
Plant material was ground to a fine powder as previously described (Horvath et al. 2019). RNA was extracted using the Qiagen RNeasy Plant Mini Kit (Cat# 74904) as per manufacturers recommendations. Two RNA samples for each tissue/treatment were randomly chosen and mixed at equal concentrations for each of the biological replicates (resulting in three biological replicates for each tissue/treatment from the six individuals collected). Biological replicate pools of total RNA were quantified and Poly A plus RNA was extracted using the New England Biolabs NEBNext Poly(A) mRNA Magnetic Isolation Module (Cat# E7490L) kit as per manufacturers instructions. Illumina sequencing libraries were constructed using the NEBNext Ultra Directional RNA Library Prep Kit for Illumina (Cat# E7760) using the suggested protocol. Tagged libraries were tested for quality using a bioanalyzer and grouped into 12-16 libraries per pool and sequenced by an outside vendor on an illumina 2500 platform with 150 base paired end reads.