A simple procedure for constructing 5'-amino-terminated oligodeoxynucleotides in aqueous solution.
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.
Bruick
R K
RK
Department of Chemistry, The Scripps Research Institute, La Jolla, CA 92037, USA.
Koppitz
M
M
Joyce
G F
GF
Orgel
L E
LE
D001483:Base Sequence; D004247:DNA / Q000138:chemical synthesis; D011088:DNA Ligases; D004591:Electrophoresis, Polyacrylamide Gel; D005022:Ethyldimethylaminopropyl Carbodiimide; D007202:Indicators and Reagents; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis* / Q000737:chemistry; D012996:Solutions; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013698:Templates, Genetic; D013936:Thymidine / Q000031:analogs & derivatives*; D014867:Water
D007202:Indicators and Reagents; D009838:Oligodeoxyribonucleotides; D012996:Solutions; D014867:Water; C023821:5-(aminomethyl)-2'-deoxyuridine; D004247:DNA; D011088:DNA Ligases; D005022:Ethyldimethylaminopropyl Carbodiimide; D013936:Thymidine