Continuous, on-line DNA sequencing using oligodeoxynucleotide primers with multiple fluorophores

J A Brumbaugh; L R Middendorf; D L Grone; J L Ruth

doi:10.1073/pnas.85.15.5610

Continuous, on-line DNA sequencing using oligodeoxynucleotide primers with multiple fluorophores

Proc Natl Acad Sci U S A. 1988 Aug;85(15):5610-4. doi: 10.1073/pnas.85.15.5610.

Authors

J A Brumbaugh¹, L R Middendorf, D L Grone, J L Ruth

Affiliation

¹ School of Biological Sciences, University of Nebraska, Lincoln 68588-0118.

Abstract

A method for sequencing DNA by using a difluoresceinated primer and laser excitation is described. Dideoxy protocols have been determined that provide sequences for 600 bases starting with base 1 with less than 1% error in a single load. Electrophoresis is at 20 W and the bands are detected 24 cm from the bottom of the loading well with a scanning fluorescence detector. Bands are imaged on a TV screen in two dimensions. The sequences can be read from the TV screen manually or semiautomatically by using a simple software program. The system allows more bases to be read with a lower error rate than any other reported automated sequencing method.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Autoradiography
Base Sequence*
Chemical Phenomena
Chemistry
DNA / genetics*
Deoxyuridine / analogs & derivatives
Electrophoresis, Polyacrylamide Gel
Fluorescent Dyes
Humans
Image Processing, Computer-Assisted
Lasers
Oligonucleotides*
Software

Substances

Fluorescent Dyes
Oligonucleotides
DNA
Deoxyuridine