Introduction to the Workshop
URLs Provided by Attendees

Abstracts

Mapping

Informatics

Sequencing

Instrumentation

Ethical, Legal, and Social Issues

Infrastructure

The electronic form of this document may be cited in the following style:
Human Genome Program, U.S. Department of Energy, DOE Human Genome Program Contractor-Grantee Workshop IV, 1994.

Abstracts scanned from text submitted for November 1994 DOE Human Genome Program Contractor-Grantee Workshop. Inaccuracies have not been corrected.

Development of Instrumentation for DNA Sequencing at a Rate of 40 Million Bases per Day

Edward S. Yeung,[1] Huan-Tsung Chang, Qingbo Li, Xiandan Lu
Ames Laboratory-USDOE and Department of Chemistry, Iowa State University, Ames, IA 50011. [1]Corresponding author.

One of the many bottlenecks in high-speed DNA sequencing using available instrumentation is the electrophoretic separation and the subsequent identification of DNA fragments derived from the Sanger reaction. A combination of fast separation by capillary electrophoresis (30 bases per minute per channel) and simultaneous monitoring (100 channels in parallel) has led to a raw sequencing rate of 3,000 bases per minute in a single instrument. This technology is readily scalable to 1,000 capillaries to achieve a gross sequencing rate of 40 million bases per day.

The substantial increase in sequencing rate is a result of several technical advances in our laboratory. (1) The use of commercial linear polymers for sieving allows replaceable yet reproducible matrices to be prepared that have lower viscosity (thus faster migration rates) compared to polyacrylamide. (2) The use of a charge-injection device camera allows random data acquisition to decrease data storage and data transfer time. (3) The use of distinct excitation wavelengths and cut-off emission filters allows maximum light throughput for efficient excitation and sensitive detection employing the standard 4-dye coding. (4) The use of index-matching and 1:1 imaging reduces stray light without sacrificing the convenience of on-column detection.