Continuous, On-line DNA Sequencing Using Oligodeoxynucleotide Primers with Multiple Fluorophores

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1988 Aug 1

PMID 3399506

Citations 29

Authors

J A Brumbaugh

L R Middendorf

D L Grone

J L Ruth

Affiliations

Soon will be listed here.

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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