Open AccessDNA Conjugation by Staudinger Ligation
Author(s) -
S. Weisbrod,
Anna Baccaro,
Andreas Marx
Publication year - 2008
Publication title -
nucleic acids symposium series
Language(s) - English
Resource type - Journals
eISSN - 1746-8272
pISSN - 0261-3166
DOI - 10.1093/nass/nrn195
Subject(s) - azide , primer (cosmetics) , ligation , chemistry , dna , staudinger reaction , primer extension , nucleotide , combinatorial chemistry , primer dimer , dna polymerase , klenow fragment , polymerase , stereochemistry , biochemistry , polymerase chain reaction , microbiology and biotechnology , biology , multiplex polymerase chain reaction , base sequence , organic chemistry , gene , exonuclease
Two 5 modified 2'-deoxyuridin triphosphates and a 7 modified 2'-deoxy-7-deazaadenosine were synthesized carrying a terminal azide linked to the base. For probing the sterical influence on incorporation and Staudinger ligation different sized flexible linkers were chosen. All three nucleotides can completely replace their natural counterparts in primer extension as well as polymerase chain reactions (PCR) using Pwo DNA polymerase. For azide labeled primer extension products subsequent conjugation of suitably functionalized phosphines via Staudinger ligation was achieved, e.g. for the conjugation of biotin as an affinity tag.
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