The real time polymerase chain reaction (rtPCR) is an essential method for detecting nucleic acids that has a wide range of clinical and research applications. Current multiplexed rtPCR is capable of detecting four to six nucleic acid targets in a single sample. However, advances in clinical medicine are driving the need to measure many more targets at once. We demonstrate a novel method which significantly increases the multiplexing capability of any existing rtPCR instrument without new hardware, software, or chemistry. The technique works by varying the relative TaqMan probe concentrations amongst targets that are measured in a single fluorometric channel. Our fluorescent amplitude modulation method generates a unique rtPCR signature for every combination of targets present in a reaction. We demonstrate this technique by measuring nine different targets across three color channels with TaqMan reporting probes, yielding a detection accuracy of 98.9% across all combinations of targets. In principle this method could be extended to measure 6 or more targets per color channel across any number of color channels without loss in specificity.
Corrugated etching techniques were used to fabricate size-tunable silicon quantum dots that luminesce under photoexcitation, tunable over the visible and near infrared. By using the fidelity of lithographic patterning and strain limited, self-terminating oxidation, uniform arrays of pillar containing stacked quantum dots as small as 2 nm were patterned. Furthermore, an array of pillars, with multiple similar sized quantum dots on each pillar, was fabricated and tested. The photoluminescence displayed a multiple, closely peaked emission spectra corresponding to quantum dots with a narrow size distribution. Similar structures can provide quantum confinement effects for future nanophotonic and nanoelectronic devices.
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