Abstract
Accurate detection of gene sequences in single cells is the ultimate challenge to polymerase chain reaction (PCR) sensitivity. Unfortunately, commonly used conventional and real-time PCR techniques are often too unreliable at that level to provide the accuracy needed for clinical diagnosis. Here we provide details of linear-after-the-exponential-PCR (LATE-PCR), a method similar to asymmetric PCR in the use of primers at different concentrations, but with novel design criteria to ensure high efficiency and specificity. Compared with conventional PCR, LATE-PCR increases the signal strength and allele discrimination capability of oligonucleotide probes such as molecular beacons and reduces variability among replicate samples. The analysis of real-time kinetics of LATEPCR signals provides a means for improving the accuracy of single cell genetic diagnosis.
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Pierce, K.E., Wangh, L.J. (2007). Linear-After-The-Exponential Polymerase Chain Reaction and Allied Technologies. In: Thornhill, A. (eds) Single Cell Diagnostics. Methods in Molecular Medicine™, vol 132. Humana Press. https://doi.org/10.1007/978-1-59745-298-4_7
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DOI: https://doi.org/10.1007/978-1-59745-298-4_7
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