Design and characterization of a gel loading mechanism for an ultra-high throughput mutational spectrometer

Abstract

A process known as Constant Denaturant Capillary Electrophoresis is used to separate mutant from wild-type DNA at fractions down to 10-7. A device known as an Ultra-high Throughput Mutational Spectrometer is being created to run 10,000 parallel channels of CDCE in order to correlate multiple point mutations in DNA with the diseases that they can cause, such as cancer. By separating the DNA in large populations, the underlying causes of such diseases can be identified. To successfully run CDCE, a high viscosity polymer gel must be loaded into each of the 10,000 channels, each of which are composed of an individual glass capillary with a 75 m inner diameter. A mechanism was designed and tested which loaded gel into 8 channels simultaneously. The mechanism was used to test the relationship between gel loading time in relation to varied pressure and capillary length, through 45 total runs, with 8 channels per run. The relationships were characterized, resulting in two equations that enable an accurate prediction of the fill time necessary to load 10,000 parallel channels simultaneously under varied conditions.