Tools

Real-Time qPCR as a Tool for Evaluating RNAi-Mediated

Real-time quantitative PCR (rt-qPCR) is the method of choice for accurate, sensitive, and specific quantitation of nucleic acid sequences. Applications of this technology are numerous in molecular diagnostics and virtually all fields of life sciences, including gene expression profiling, measurement
of DNA copy number alterations, genotyping, mutation detection, pathogen detection, measurement of viral load, disease monitoring, and assessment of drug response. Several factors, such as careful primer design and evaluation,
template preparation, implementation of a robust normalization strategy, and accurate data analysis, are essential for a successful and reliable rt-qPCR assay. In this report we describe how rt-qPCR can be implemented as a tool for monitoring silencing efficiency and assessing functional effects of RNA interference (RNAi)-mediated knockdown of MYCN and TP53 genes, which are pivotal in neuroblastoma pathogenesis.

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