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Gene expression comprises multiple cellular processes, from transcription, processing, and export of mRNA from the nucleus to translation into protein and post-translational modifications. Each stage is tightly regulated by the cell, and overall expression levels reflect the sum of these steps. Molecular techniques are commonly used for assessing gene expression because they are fast, sensitive, and relatively inexpensive. mRNA quantification exploits antisense hybridization either directly, as in the case of microarray, RNase protection assay, and northern blotting, or with subsequent amplification, such as polymerase chain reaction (PCR).
Digital PCR (dPCR) is a valuable tool to complement other molecular techniques across a wide range of applications, including gene expression analysis. dPCR divides the reaction into many smaller micro-reactions, each of which is read individually and interpreted as positive or negative based on the presence or absence of fluorescence (endpoint detection).
Separating the targets into micro-reactions permits direct quantification of the target sequence (through Poisson statistics) without the need for reference standards or a standard curve. Digital PCR excels at providing highly precise quantification, even at low concentrations, and can often detect small differences more reliably than other techniques. Because it relies on endpoint detection, dPCR results are not impacted by variations in PCR efficiency and is generally more robust and reliable for working with challenging samples that contain PCR inhibitors or low-abundance targets.
Explore more details about gene expression analysis by dPCR in our technical note.
For Research Use Only. Not for use in diagnostic procedures.