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Frequently asked questions
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.
How can I assess knockdown of my target gene?
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.
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