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PMID: 2668874 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Direct solid phase sequencing of genomic and plasmid DNA using magnetic beads as solid support.

Nucleic acids research ·Vol. 17 ·No. 13 ·1989-07-11 ·Pages 4937-46

Hultman T, Ståhl S, Hornes E, Uhlén M

Abstract

Approaches to direct solid phase sequencing of genomic and plasmid DNA have been developed using magnetic beads, coated with streptavidin, as solid support. The DNA is immobilized through selective incorporation of biotin into one of the strands. A single stranded template, suitable for sequencing, is obtained through strand-specific elution. Using this concept, in vitro amplified plasmid DNA and chromosomal DNA were sequenced directly from single colonies. The solid phase approach ensures that the amplification and the sequencing reactions can be performed under optimal conditions. The system was found to be suitable for sequencing using both isotope- and fluorescent-labelled primers.

MeSH Terms
Base Sequence DNA, Bacterial/genetics,isolation & purification DNA-Directed DNA Polymerase Enterotoxins/genetics Escherichia coli/genetics Gene Amplification Genes, Bacterial Genetic Techniques Molecular Sequence Data Plasmids Staphylococcus aureus/genetics Templates, Genetic
Chemicals
DNA, Bacterial Enterotoxins enterotoxin A, Staphylococcal DNA-Directed DNA Polymerase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Hultman T
Department of Biochemistry and Biotechnology, Royal Institute of Technology, Stockholm, Sweden.
Ståhl S
Hornes E
Uhlén M
References (18)
18 references, click to expand
  1. Host controlled restriction mutants of Staphylococcus aureus.
    Arch Roum Pathol Exp Microbiol. 1975 Jan-Jun;34(1-2):55-8 PMID: 1212076
  2. Automation of dideoxynucleotide DNA sequencing reactions using a robotic workstation.
    Biotechniques. 1988 Sep;6(8):776-7, 781-7 PMID: 3273640
  3. Uniformly spaced banding pattern in DNA sequencing gels by use of field-strength gradient.
    J Biochem Biophys Methods. 1984 Nov;10(1-2):83-90 PMID: 6520329
  4. Fluorescence detection in automated DNA sequence analysis.
    Nature. 1986 Jun 12-18;321(6071):674-9 PMID: 3713851
  5. Direct cloning and sequence analysis of enzymatically amplified genomic sequences.
    Science. 1986 Sep 5;233(4768):1076-8 PMID: 3461561
  6. Length mutations in human mitochondrial DNA: direct sequencing of enzymatically amplified DNA.
    Nucleic Acids Res. 1987 Jan 26;15(2):529-42 PMID: 2881260
  7. Automated DNA sequencing: ultrasensitive detection of fluorescent bands during electrophoresis.
    Nucleic Acids Res. 1987 Jun 11;15(11):4593-602 PMID: 3588303
  8. A system for rapid DNA sequencing with fluorescent chain-terminating dideoxynucleotides.
    Science. 1987 Oct 16;238(4825):336-41 PMID: 2443975
  9. Characterization of beta-thalassaemia mutations using direct genomic sequencing of amplified single copy DNA.
    Nature. 1987 Nov 26-Dec 2;330(6146):384-6 PMID: 3683554
  10. Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction.
    Methods Enzymol. 1987;155:335-50 PMID: 3431465
  11. Genomic amplification with transcript sequencing.
    Science. 1988 Jan 29;239(4839):491-4 PMID: 3340835
  12. Solid phase DNA sequencing using the biotin-avidin system.
    Nucleic Acids Res. 1988 Apr 11;16(7):3025-38 PMID: 3368314
  13. T7 DNA polymerase in automated dideoxy sequencing.
    Nucleic Acids Res. 1988 Apr 25;16(8):3487-96 PMID: 3375061
  14. Automated Sanger dideoxy sequencing reaction protocol.
    FEBS Lett. 1988 Jun 20;233(2):432-6 PMID: 3289973
  15. Generation of single-stranded DNA by the polymerase chain reaction and its application to direct sequencing of the HLA-DQA locus.
    Proc Natl Acad Sci U S A. 1988 Oct;85(20):7652-6 PMID: 3174659
  16. DNA sequencing with Thermus aquaticus DNA polymerase and direct sequencing of polymerase chain reaction-amplified DNA.
    Proc Natl Acad Sci U S A. 1988 Dec;85(24):9436-40 PMID: 3200828
  17. A new method of sequencing DNA.
    Anal Biochem. 1988 Nov 1;174(2):423-36 PMID: 2853582
  18. Complete sequence of the staphylococcal gene encoding protein A. A gene evolved through multiple duplications.
    J Biol Chem. 1984 Feb 10;259(3):1695-702 PMID: 6319407
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
Published
1989-07-11
Pages
4937-46
Language
English
Region
England
NLM ID
0411011
PMCID
PMC318085
Subset
IM
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