Synthetic DNA templates for the production of in situ hybridization probes

Abstract

Generating RNA riboprobes for in situ hybridization generally requires the use of plasmids, which must be grown in bacteria, isolated, purified, and linearized prior to in vitro transcription. Here we report a simple method for generating DNA templates for the in vitro transcription of RNA probes from synthetic DNA (IDT gBlocks). Each synthetic DNA template contains sequences corresponding to the target mRNA flanked by bacteriophage promoters. Amplification of the template by a single round of PCR and subsequent in vitro transcription results in production of high quality RNA probes for in situ hybridization.