A simple procedure for constructing 5'-amino-terminated oligodeoxynucleotides in aqueous solution

R K Bruick; M Koppitz; G F Joyce; L E Orgel

doi:10.1093/nar/25.6.1309

A simple procedure for constructing 5'-amino-terminated oligodeoxynucleotides in aqueous solution

Nucleic Acids Res. 1997 Mar 15;25(6):1309-10. doi: 10.1093/nar/25.6.1309.

Authors

R K Bruick¹, M Koppitz, G F Joyce, L E Orgel

Collaborator

S L Miller²

Affiliations

¹ Department of Chemistry, The Scripps Research Institute, La Jolla, CA 92037, USA.
² U CA, San Diego, La Jolla

Abstract

A rapid method for the synthesis of oligodeoxynucleotides (ODNs) terminated by 5'-amino-5'-deoxythymidine is described. A 3'-phosphorylated ODN (the donor) is incubated in aqueous solution with 5'-amino- 5'-deoxythymidine in the presence of N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDC), extending the donor by one residue via a phosphoramidate bond. Template- directed ligation of the extended donor and an acceptor ODN, followed by acid hydrolysis, yields the acceptor ODN extended by a single 5'-amino-5'-deoxythymidine residue at its 5'terminus.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Base Sequence
DNA / chemical synthesis
DNA Ligases
Electrophoresis, Polyacrylamide Gel
Ethyldimethylaminopropyl Carbodiimide
Indicators and Reagents
Oligodeoxyribonucleotides / chemical synthesis*
Oligodeoxyribonucleotides / chemistry
Solutions
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Templates, Genetic
Thymidine / analogs & derivatives*
Water

Substances

Indicators and Reagents
Oligodeoxyribonucleotides
Solutions
Water
5-(aminomethyl)-2'-deoxyuridine
DNA
DNA Ligases
Ethyldimethylaminopropyl Carbodiimide
Thymidine