Measurement of mRNA Expression Knockdown Using siRNA Through RT-qPCR
Description
This template was adapted from the original submission. Edits were made to enhance scientific accuracy, optimal usability and/or to meet industry-leading design standards for science communication.
This diagram highlights the general workflow for validating siRNA knockdown efficiency for a target gene and determining mRNA expression for other genes of interest. First, siRNA is transfected into cells and incubated for 48-72 hours for efficient knockdown. Next, RNA is either isolated following reverse transcription to cDNA, or stored at -80 °C for future use. Lastly, the cDNA is amplified through qPCR and mRNA is assessed through corresponding Ct values.
Acknowledgements
References
Tuzmen, S.. (2007) Validation of Short Interfering RNA Knockdowns by Quantitative Real-Time PCR. Methods Mol Biol.. https://doi.org/10.1385/1-59745-229-7:177
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