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PMID: 3368314 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Solid phase DNA sequencing using the biotin-avidin system.

Nucleic acids research ·Vol. 16 ·No. 7 ·1988-04-11 ·Pages 3025-38

Ståhl S, Hultman T, Olsson A, Moks T, Uhlén M

Abstract

A novel method for solid-phase DNA sequencing is described. A plasmid vector, pRIT27, has been designed to allow directional immobilization of double stranded plasmid to avidin agarose. The strategy involves enzymatic incorporation of 11-bio-dUTP into the plasmid and strand specific elution using alkali. The immobilized single stranded DNA is used as template for sequencing reactions and the resulting labelled oligonucleotides are eluted by alkali. The affinity gel containing the immobilized template is consecutively used for the four different dideoxy-nucleotide reactions. The solid-phase technique can be used for both primer specific or extension specific labelling. The possibility to use the system in automated DNA sequencing is discussed.

MeSH Terms

Autoanalysis/methods Avidin Base Sequence Biotin Cloning, Molecular/methods DNA/isolation & purification Deoxyribonucleotides Enzymes, Immobilized Genetic Vectors Plasmids Sepharose Sodium Hydroxide

Chemicals

Deoxyribonucleotides Enzymes, Immobilized Avidin Sodium Hydroxide Biotin DNA Sepharose

Authors & Affiliations

5 authors, click to expand affiliations / ORCID

Ståhl S

Department of Biochemistry, Royal Institute of Technology, Stockholm, Sweden.

Hultman T

Olsson A

Moks T

Uhlén M

References (14)

14 references, click to expand

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Article Info

Journal

Nucleic acids research

Abbr.

Nucleic Acids Res

ISSN

0305-1048

DOI

10.1093/nar/16.7.3025

Published

1988-04-11

Pages

3025-38

Language

English

Region

England

NLM ID

0411011

PMCID

PMC336449

Subset

External Links

PubMed source DOI: 10.1093/nar/16.7.3025 PMC Full Text View journal details

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