Abstract
The following protocols provide a rapid approach for establishing good working conditions for transfecting siRNAs for specific gene knockdown. By first using microscopy to evaluate efficient transfection of an inexpensive, fluorescent oligonucleotide, the researcher can later proceed with more expensive Western blot or quantitative real-time PCR (qRT-PCR) methods. Thus, the main culprit of ineffective knockdown, poor transfection, can be eliminated before engaging in tedious and time-consuming approaches for troubleshooting siRNA knockdown experiments.
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Acknowledgments
This work is generously supported by funding from the Singapore Agency for Science, Technology and Research (A*STAR).
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Chen, S.W., Oh, S.K. (2010). Establishing Efficient siRNA Knockdown in Stem Cells Using Fluorescent Oligonucleotides. In: Zhang, B., Stellwag, E. (eds) RNAi and microRNA-Mediated Gene Regulation in Stem Cells. Methods in Molecular Biology, vol 650. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-769-3_6
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DOI: https://doi.org/10.1007/978-1-60761-769-3_6
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Publisher Name: Humana Press, Totowa, NJ
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