Oligonucleotide dendrimers: synthesis and use as polylabelled DNA probes.
Oligonucleotide dendrimers were synthesized using a novel phosphoramidite synthon, tris-2,2,2-[3-(4,4'-dimethoxytrityloxy) propyloxymethyl]ethyl- N , N -diisopropylaminocyanethoxy phosphoramidite. Label, incorporated using [gamma-32P]ATP and polynucleotide kinase, was increased in proportion to the number of 5'-ends. There was a similar increase in signal when these multiply labelled oligonucleotides were used as probes to oligonucleotide arrays. A dendrimeric oligonucleotide was used successfully as a primer in the PCR. The strand bearing the dendrimer was resistant to degradation by T7 Gene 6 exonuclease making it easy to convert the double-stranded product of the PCR to a multiply-labelled, single-stranded probe.
Shchepinov
M S
MS
Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK. misha@bioch.ox.ac.uk
Udalova
I A
IA
Bridgman
A J
AJ
Southern
E M
EM
D000255:Adenosine Triphosphate; D001704:Biopolymers; D017931:DNA Primers / Q000138:chemical synthesis*; D015342:DNA Probes / Q000138:chemical synthesis*; D007553:Isotope Labeling; D009841:Oligonucleotides / Q000138:chemical synthesis*; D010761:Phosphorus Radioisotopes; D016133:Polymerase Chain Reaction; D011116:Polynucleotide 5'-Hydroxyl-Kinase
D001704:Biopolymers; D017931:DNA Primers; D015342:DNA Probes; D009841:Oligonucleotides; D010761:Phosphorus Radioisotopes; D000255:Adenosine Triphosphate; D011116:Polynucleotide 5'-Hydroxyl-Kinase