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Journal: International Journal of Molecular Sciences
Article Title: The Potential and Limitations of the MinION/Yenos Platform for miRNA-Enabled Early Cancer Detection
doi: 10.3390/ijms26083822
Figure Lengend Snippet: From [ , , ]. A graphical abstract of the processes involved in the miRNA measurement using the MinION/Yenos platform. Left, top: Collection of the biospecimen, blood, or urine, followed by total RNA isolation using a commercial kit and measurement of total RNA in the isolate using a nanodrop spectrophotometer. Left, bottom: Mixing an aliquot from the RNA isolate with an aliquot of the probe complementary to the target miRNA (Materials), adding ONT buffer and conducting a MinION unassisted ion conductance experiment (two experiments running simultaneously, shown here). The experiment measures the ion current ( I ) in picoamperes (pA) as a function of time (t) in milliseconds (ms). I is a constant at Io , which is the open nanopore ion current ( Io ). When a single molecule traverses the nanopore, Io is reduced to a new value, Ir , because the molecule occupies the space that would have been occupied by the electrolyte that produces Io . Ion current reduction (dip in this platform) lasts for a time, τ, and produces an event that is read by software ( OsBp_detect , see ). The analysis determines whether the free probe is in excess and detected (left on the scheme above) or if the probe is not detected because it is hybridized with the target (right on the scheme above). Notably, RNAs, including the target miRNA, traverse much faster than the probes, and they are mostly missed (bottom on the scheme above) due to the relatively slow acquisition rate of this platform. A probe is an oligodeoxynucleotide complementary to the target miRNA, loaded with osmium tags (see , Yenos probes), optimized for hybridization with the target and for detection by the nanopores .
Article Snippet: For the isolation of total
Techniques: Isolation, Spectrophotometry, Software, Hybridization