Fast DNA Sequencing Research Articles

Fast DNA sequencing

A method for fast nucleotide sequencing is described. It is based in the selection of well-known small oligomers able to be hybridized with the unknown target. The selected oligomers are afterwards ordered following a simple statistical approach. The use of capillary electrophoresis for the analysis is emphasized.

Ultrafast DNA analysis by capillary electrophoresis/laser-induced fluorescence detection.

The limits of ultrafast DNA analysis by CE were determined by investigating the influence of the effective capillary length and the electric field strength on the analysis time for a given peak resolution (10 bp). In accordance with theory, the use of a fast ramp power supply for narrow plug electrokinetic injection was found to be essential to minimize the extra column effects on peak dispersion. Two major column dispersion factors, longitudinal diffusion and thermal dispersion, were determined experimentally, as well as the influence of the electric field strength on the electrophoretic mobilities and diffusion coefficients of DNA. It was found that higher field strengths can be applied with lower thermal dispersion than predicted by classical CE models. This was attributed to the faster mass transport in the radial direction due to field-induced DNA orientation. Short capillaries (approximately 3-7 cm effective length) and moderate to high electric field strengths (approximately 600-800 V/cm) were used to perform a series of fast DNA separations. The dsDNA fragment standards phiX174/HaeIII and pBR322/HaeIII were separated within 30 s. The possibility for fast mutation detection was demonstrated using constant denaturant capillary electrophoresis (CDCE) for the analysis of a single base mutation in mitochondrial DNA in 72 s. The potential for fast DNA sequencing was illustrated by separating 300 ssDNA fragments within 180 s.

Fast plasmid DNA sequencing using a thermal cycler and high temperature alkali denaturation.

Fast plasmid DNA sequencing using a thermal cycler and high temperature alkali denaturation.

DNA sequencing by mass spectrometry.

Mass spectrometry has the potential to replace gel electrophoresis for fast DNA sequencing. In this tutorial paper, it is discussed how far mass spectrometry of DNA has come since the advent of electrospray ionization (ESI) and matrix-assisted laser desorption/ionization (MALDI) and how much remains to be done before mass spectrometry will be a useful tool for DNA sequencing. A brief description of MALDI and ESI mass spectrometric analysis of DNA is presented along with the different strategies for DNA sequencing using mass spectrometry. These include mass spectrometric analysis to replace gel separation in Sanger sequencing, enzymatic ladder sequencing and sequencing by gas-phase fragmentation. The future role of mass spectrometry in DNA sequencing in the Human Genome Project and beyond is also discussed.

MALDI for fast DNA analysis and sequencing

Matrix-assisted laser desorption/ionization (MALDI) has been pursued for fast DNA analysis and sequencing. In this article, recent progress on detecting large oligonucleotides is reviewed. The potential application of MALDI for population screening for disease diagnosis is discussed. The approach of using MALDI for DNA sequencing is presented. Discussion on the use of robotic and automation to speed up the mass spectrometer sequencing process is also included. © 1996 John Wiley & Sons, Inc.

MALDI for fast DNA analysis and sequencing

MALDI for fast DNA analysis and sequencing

Fast DNA sequencing technique developed

Chemists at the University of Wisconsin, Madison, have developed an approach for significantly increasing the rate at which the fragments produced in DNA sequencing reactions can be analyzed by gel electrophoresis. The research was described at a symposium sponsored by the Biotechnology Secretariat. The technique, which utilizes ultrathin gels, provides more than 9000 bases of DNA sequence data per hour. Current commercial sequencing instruments produce about 1350 bases of DNA sequence data per hour. And Lloyd M. Smith, an assistant chemistry professor at Wisconsin who developed the technique, says that fairly straightforward extensions of the current design will enable construction of a system with a throughput of 26,000 bases per hour. Smith points out that high-speed DNA sequencing is an absolute necessity if the human genome project and related sequencing efforts are to succeed. To date, about 50 million base pairs of DNA sequence have been published, about 6 million of which are from the ...

A simplified protocol for fast plasmid DNA sequencing

A simplified protocol for fast plasmid DNA sequencing