Improved high-throughput DNA fragment analyzer employing horizontal ultrathin gel electrophoresis

Robert L. Brumley Jr.; John A. Luckey

doi:10.1117/12.237623

1 April 1996 Improved high-throughput DNA fragment analyzer employing horizontal ultrathin gel electrophoresis

Robert L. Brumley Jr., John A. Luckey

Proceedings Volume 2680, Ultrasensitive Biochemical Diagnostics; (1996) https://doi.org/10.1117/12.237623
Event: Photonics West '96, 1996, San Jose, CA, United States

Abstract

We are currently developing a significantly improved gel electrophoresis and detection system that will allow more than an order of magnitude enhancement in the speed of DNA fragment analysis. This system is based upon the technique of horizontal ultrathin gel electrophoresis (HUGE) which employs denaturing polyacrylamide gels that are 75 microns thick. Because of the thinness of the gel, very high electric field strengths may be applied without deleterious thermal effects on resolution. Our proprietary fluorescence detector that scans the gel during electrophoresis allows for the simultaneous detection of up to four fluorophores. Because of the efficiency of the system of light collection, the gel can be scanned at speeds fast enough to generate high resolution gel images despite the high speed of separations. In addition, we are able to increase sample density by collecting 500 datapoints across the width of the gel. The resulting instrument has the capability to separate and resolve single-stranded DNA molecules that are between 25 and 300 bases in length from each of 60 lanes in less than 45 minutes. With the advent of 96 lane gels and attendant automation, this instrument will have the ability to analyze 18,432 genotypes per day.

Citation Download Citation

Robert L. Brumley Jr. and John A. Luckey "Improved high-throughput DNA fragment analyzer employing horizontal ultrathin gel electrophoresis", Proc. SPIE 2680, Ultrasensitive Biochemical Diagnostics, (1 April 1996); https://doi.org/10.1117/12.237623

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