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PMID: 4000959 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The synthesis of oligonucleotides containing an aliphatic amino group at the 5' terminus: synthesis of fluorescent DNA primers for use in DNA sequence analysis.

Nucleic acids research ·Vol. 13 ·No. 7 ·1985-04-11 ·Pages 2399-412

Smith LM, Fung S, Hunkapiller MW, Hunkapiller TJ, Hood LE

Abstract

A rapid and versatile method has been developed for the synthesis of oligonucleotides which contain an aliphatic amino group at their 5' terminus. This amino group reacts specifically with a variety of electrophiles, thereby allowing other chemical species to be attached to the oligonucleotide. This chemistry has been utilized to synthesize several fluorescent derivatives of an oligonucleotide primer used in DNA sequence analysis by the dideoxy (enzymatic) method. The modified primers are highly fluorescent and retain their ability to specifically prime DNA synthesis. The use of these fluorescent primers in DNA sequence analysis will enable DNA sequence analysis to be automated.

MeSH Terms
Base Sequence Chemical Phenomena Chemistry Chromatography, High Pressure Liquid DNA/analysis Fluorescent Dyes Methods Oligonucleotides/chemical synthesis
Chemicals
Fluorescent Dyes Oligonucleotides DNA
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Smith L M
Fung S
Hunkapiller M W
Hunkapiller T J
Hood L E
References (7)
7 references, click to expand
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  4. In situ activation of bis-dialkylaminophosphines--a new method for synthesizing deoxyoligonucleotides on polymer supports.
    Nucleic Acids Res. 1984 May 25;12(10):4051-61 PMID: 6728676
  5. Derivatization of unprotected polynucleotides.
    Nucleic Acids Res. 1983 Sep 24;11(18):6513-29 PMID: 6622259
  6. Synthesis of oligonucleotides containing the hypermodified base, alpha-putrescinylthymine.
    Nucleic Acids Symp Ser. 1983;(12):75-8 PMID: 6664879
  7. Sequencing end-labeled DNA with base-specific chemical cleavages.
    Methods Enzymol. 1980;65(1):499-560 PMID: 6246368
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
Published
1985-04-11
Pages
2399-412
Language
English
Region
England
NLM ID
0411011
PMCID
PMC341163
Subset
IM
Grants
NIGMS NIH HHS · GM07401 · United States
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