High-Throughput Targeted Transcriptional Profiling of Defense Genes Using RNA-Mediated Oligonucleotide Annealing, Selection, and Ligation with Next-Generation Sequencing in Arabidopsis.

Tracking RNA transcription has been one of the most powerful tools to gain insight into the biological process. While a wide range of molecular methods such as northern blotting, RNA-seq, and quantitative RT-PCR are available, one of the barriers in transcript analysis is an inability to accommodate a sufficient number of samples to achieve high resolution in dynamic transcriptional changes. RASL-seq (RNA-mediated oligonucleotide Annealing, Selection, and Ligation with next-generation sequencing) is a sequencing-based transcription profiling tool that processes hundreds of samples assessing a set of over a hundred genes with a fraction of the cost of a conventional RNA-seq. We described a RASL-seq protocol for assessing 288 genes mostly including defense genes to capture their dynamic nature. We demonstrated that this transcriptional profiling method produced a highly reliable outcome comparable to a conventional RNA-seq and quantitative RT-PCR.