DNA mutation detection via fluorescence imaging in a spatial thermal gradient, capillary electrophoresis system

Abstract

To improve the speed and efficiency of genetic screening, we are developing an instrument to detect mutations via a high throughput, automated system. Detection is based on changes in the melting temperature induced by single point mutations as in Denaturing Gradient Gel Electrophoresis (DGGE). This instrument measures the migrating position of test and wild type molecules in a spatial thermal gradient during capillary electrophoresis. In this thesis, the concept is described. A thorough design process is conducted focusing on controlling thermal expansion, creating a stable, predictable temperature gradient, and optimizing software and hardware. Finally, the concept is proved in a series of experiments successfully identifying the presence of a mutation in the test sample.

DNA Diagnostics by Capillary Electrophoresis

“…649 Several papers focused on development of high-throughput automated instruments with precise control of spatial and temporal temperature gradients. 650 Yeung's group used two simple heating devices controlling a temperature gradient in separation capillaries for detection of unknown point mutations. A temporal gradient was established by the temperature programming of an aluminum heating plate connected to an array of 96 capillaries through a thermal conductive paste.…”

“…Several papers focused on development of high-throughput automated instruments with precise control of spatial and temporal temperature gradients . Yeung's group used two simple heating devices controlling a temperature gradient in separation capillaries for detection of unknown point mutations.…”

Automated analysis of behavior: A computer‐controlled system for drug screening and the investigation of learning

“…Ultimately, massively scaled-up and miniaturized versions of this device will be used for highthroughput screens of novel neuroactive compounds (Kanigan et al, 2000;Crane et al, 2001;Anquetil et al, 2003;Martel and Hunter, 2006). Through discovery of such reagents, and of endogenous genetic components regulating aspects of behavior, we envision new vistas in cognitive science.…”

A multiscale products technique for denoising of DNA capillary electrophoresis signals

“…Capillary electrophoresis (CE) [1,2] is a well-developed technology and is widely used in chemistry and biomedical application. Due to high sensitivity, small injection volumes and reduced consumption of solvent and samples, CE is also used for sequencing and separation of high-throughput DNA [3]. However, CE signals are often corrupted by random noise resulting from the variation of instrument and the physicalchemical processes of separation.…”