One hundred channel electrophoresis prototypes for application to an ultra-high throughput mutational spectrometer

Abstract

Mutation detection within the human genome is becoming an increasingly important field today. It is possible that with a correlation between diseases and the mutations that cause them, new therapeutic treatments could be developed against many of today's common diseases In order to accomplish mutation detection, 1012 gene segments may be needed, requiring a significant increase in current technologies An instrument termed an Ultra-high throughput mutational spectrometer (UTMS) uses a process known as constant denaturing capillary electrophoresis (CDCE) to detect mutations in 10,000 capillaries simultaneously. The UTMS is at a 100-capillary proof of concept stage to successfully perform CDCE. In order for this stage to be successful, multiple subsystems of the device must work in unison, including thermal control, optical detection, electrical and fluidic connectivity. In this thesis, multiple devices were created to work in conjunction with previously existing instruments, including a passively aligned 100-port buffer reservoir and DNA injection loading plate. These devices were used to perform electrophoresis on DNA fluorescent primers in order to test the functionality of the UTMS 100-stage concept.

(cont.) A procedure for performing these experiments was developed in order to minimize risk and maximize chance of success. Successful isolation of individual signal detection was accomplished through the use of these procedures and devices, proving the 100-capillary proof of concept may reliably and repeatedly perform CDCE on the UTMS.