Visualization of RNA Transcripts in Western Corn Rootworm (Diabrotica virgifera virgifera) and Plants by In Situ Hybridization.

In situ hybridization (ISH) is a methodology by which nucleic acids are detected within fixed tissue samples. Recent advances in detection technology and target recovery have greatly enhanced the technique's ability to detect single mRNA molecules. Here we detail the fixation, paraffin embedding, sectioning, target recovery, and chromogenic detection of an mRNA (DvSSJ1), encoding for a membrane protein associated with the smooth septate junction (SSJ) in Western corn rootworm [Diabrotica virgifera (Dv)]. Further, we demonstrate, the expression of dsRNA of DvSSJ1 in maize root tissues using signal amplification and background suppression technology.