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PMID: 3399506 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

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

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 85 ·No. 15 ·1988-08-00 ·Pages 5610-4

Brumbaugh JA, Middendorf LR, Grone DL, Ruth JL

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.

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
Chemicals
Fluorescent Dyes Oligonucleotides DNA Deoxyuridine
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Brumbaugh J A
School of Biological Sciences, University of Nebraska, Lincoln 68588-0118.
Middendorf L R
Grone D L
Ruth J L
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17 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.85.15.5610
Published
1988-08-00
Pages
5610-4
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC281809
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.85.15.5610 PMC Full Text View journal details
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