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. 1988 Aug;85(15):5610-4.
doi: 10.1073/pnas.85.15.5610.

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

J A Brumbaugh  1 , L R Middendorf, D L Grone, J L Ruth
Affiliations

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

J A Brumbaugh et al. Proc Natl Acad Sci U S A. 1988 Aug.

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.

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