3 research outputs found
High-throughput binding characterization of RNA aptamer selections using a microplate-based multiplex microcolumn device
Microfluidic Device for Aptamer-Based Cancer Cell Capture and Genetic Mutation Detection
We present a microfluidic
device for specifically capturing cancer
cells and isolating their genomic DNA (gDNA) for specific amplification
and sequence analysis. To capture cancer cells within the device,
nucleic acid aptamers that specifically bind to cancer cells were
immobilized within a channel containing micropillars designed to increase
capture efficiency. The captured cells were lysed <i>in situ</i>, and their gDNA was isolated by physical entanglement within a second
smaller-dimensioned micropillar array. This type of isolation allows
the gDNA to be retained and purified within the channel and enables
amplification and analysis to be performed on the gDNA without the
loss of the original template. We developed a technique for selectively
amplifying genes from whole gDNA using multiple displacement amplification.
The amplified gene samples were sequenced, and the resulting sequence
information was compared against the known wild-type gene to identify
any mutations. We have tested cervical and ovarian cancer cells for
mutations in the <i>TP53</i> gene using this technology.
This approach offers a way to monitor multiple genetic mutations in
the same small population of cells, which is beneficial given the
wide diversity in cancer cells, and therefore it requires very few
cells to be extracted from a patient sample