Root transcriptome RNA-Seq of Parasponia andersonii: C46 empty vector Nodulation permissive replicate 3
Root samples from each treatment, weighing between 50-70 mg, were subjected to tissue lysis using a Qiagen TissueLyser LT. This was followed by sequential phenol:chloroform and chloroform extractions. RNA was precipitated using isopropanol, washed with ethanol, and treated with DNase to eliminate genomic DNA. After a secondary phenol:chloroform and chloroform extraction, RNA was re-precipitated in ethanol and re-dissolved in nuclease-free water. RNA concentration was ascertained using both a Biochrom SimpliNano Spectrophotometer and a Qubit 2.0 Fluorometer with Broad Range Assay Kits. RNA integrity was verified by gel electrophoresis. Quantitative PCR assessed NSP2 expression, with Actin and Elongation Factor Alpha 1 as normalization references.