5' end fluorescent labelling of oligonucleotides with riboflavin-derived phosphitylating reagent.
Riboflavin was transformed within six steps into 3-isobutyryl-7,8-dimethyl-10-[2-O-(beta-cyanoethoxy-N,N- diisopropylaminophosphinyl)ethyl]isoalloxazine. This new fluorescent reagent was applied for direct phosphitylation of 5-OH function of protected oligonucleotide assembled on controlled-pore glass support by beta-cyanoethyl phosphoramidite chemistry. As the result of subsequent P(III)----P(V) oxidation and removal of protecting groups with concentrated ammonia, an oligonucleotide 5-labelled with fluorescent flavin moiety could be obtained. Using this procedure 15-mer oligonucleotide of a sequence corresponding to M13 hybridization primer was prepared.
Mielewczyk
S
S
Institute of Bioorganic Chemistry, Polish Academy of Sciences, Poznań, Poland.
Wieckowska
D
D
Krzymańska-Olejnik
E
E
Gdaniec
Z
Z
Adamiak
R W
RW
D001483:Base Sequence; D005456:Fluorescent Dyes / Q000138:chemical synthesis* / Q000737:chemistry; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000032:analysis* / Q000138:chemical synthesis / Q000737:chemistry
D005456:Fluorescent Dyes; D009841:Oligonucleotides