Our Scientific Applications Support team has assembled a list of frequently asked questions to help you find answers quickly. Filter using one or more categories to focus on specific topics, or use the search bar to perform a text search.
IDT recommends using reverse transcription quantitative PCR (RT-qPCR) to assess knockdown of your RNA target. The IDT PrimeTime™ Predesigned qPCR Primer and Probe Sets or a custom PrimeTime qPCR Assay can be used to assess gene expression for human, mouse, and rat transcriptomes.
We recommend at least two qPCR assays designed a distance apart from each other on the transcript to help control for artifacts. For example, you may get a false negative result if there are gene isoforms that are not targeted by the ASO but are still quantified by the qPCR reaction. Alternatively, you may get a false positive result if the ASO blocks the reverse transcriptase enzyme during cDNA synthesis, thus preventing full-length RNA transcript conversion to cDNA. Having concordant data for multiple qPCR assays can help increase confidence in your results.
*RUO—For research use only. Not for use in diagnostic procedures. Unless otherwise agreed to in writing, IDT does not intend for these products to be used in clinical applications and does not warrant their fitness or suitability for any clinical diagnostic use. Purchaser is solely responsible for all decisions regarding the use of these products and any associated regulatory or legal obligations.